Benzimidazole derivatives as PI3 kinase inhibitors

ABSTRACT

This invention relates to the use of benzimidazole derivatives for the modulation, notably the inhibition of the activity or function of the phosphoinositide 3′ OH kinase family (hereinafter PI3 kinases), suitably, PI3Kα, PI3Kδ, PI3Kβ, and/or PI3Kγ. Suitably, the present invention relates to the use of benzimidazoles in the treatment of one or more disease states selected from: autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, cancer, sperm motility, transplantation rejection, graft rejection and lung injuries. More suitably, the present invention relates to PI3Kβ selective benzimidazoles compounds for treating cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is filed as a continuation application of U.S. Ser. No.14/031,136 filed on Sep. 19, 2013, which is a continuation of U.S. Ser.No. 13/876,853 filed on Mar. 29, 2013 (now abandoned), which was filedpursuant to 35 USC 371 as a United States National Phase Application ofInternational Patent Application Serial No. PCT/US2011/052857 filed onSep. 23, 2011, which claims priority from 61/390,314 filed on Oct. 6,2010 and 61/528,397 filed on Aug. 29, 2011 in the United States.

FIELD OF THE INVENTION

This invention relates to the use of benzimidazole derivatives for themodulation, notably the inhibition of the activity or function of thephosphoinositide 3′ OH kinase family (hereinafter PI3 kinases),suitably, PI3Kα, PI3Kδ, PI3Kβ, and/or PI3Kγ. Suitably, the presentinvention relates to the use of benzimidazoles in the treatment of oneor more disease states selected from: autoimmune disorders, inflammatorydiseases, cardiovascular diseases, neurodegenerative diseases, allergy,asthma, pancreatitis, multiorgan failure, kidney diseases, plateletaggregation, cancer, sperm motility, transplantation rejection, graftrejection and lung injuries. More suitably, the present inventionrelates to PI3Kβ selective benzimidazoles compounds for treating cancer.

BACKGROUND OF THE INVENTION

The phosphoinositide 3-kinase (PI3K) pathway is among the most commonlyactivated in human cancer and the importance in carcinogenesis is wellestablished (Samuels Y and Ericson K. Oncogenic PI3K and its role incancer. Current Opinion in Oncology, 2006; 18:77-82). Initiation ofsignaling begins with the phosphorylation ofphosphatidylinositol-4,5-bisphosphate (PIP2) to producephosphatidylinositol-3,4,5-P3 (PIP3). PIP3 is a critical secondmessenger which recruits proteins that contain pleckstrin homologydomains to the cell membrane where they are activated. The most studiedof these proteins is AKT which promotes cell survival, growth, andproliferation.

The PI3K family consists of 15 proteins that share sequence homology,particularly within their kinase domains, but have distinct substratespecificities and modes of regulation (Vivanco I and Sawyers C L. Thephosphatidylinositol 3-kinase-AKT pathway in human cancer. NatureReviews Cancer, 2002; 2:489-501). Class I PI3Ks are heterodimersconsisting of a p110 catalytic subunit complexed to one of severalregulatory subunits collectively referred to as p85 and have been themost extensively studied in the context of tumorgenesis. The class 1API3K catalytic subunits comprise the p110α, p110β, and p110δ isoforms,which associate with one of five different regulatory subunits encodedby three separate genes. A single class 1B PI3K catalytic isoform p110γinteracts with one of two associated regulatory subunits (Crabbe T,Welham M J, Ward S G, The PI3k inhibitor arsenal: choose your weaponTrends in Biochem Sci, 2007; 32:450-456). Class 1 PI3Ks are primarilyresponsible for phosphorylating the critical PIP2 signaling molecule.

The link between the PI3K pathway and cancer was confirmed by a studywhich identified somatic mutations in the PIK3CA gene encoding the p110αprotein. Subsequently, mutations in PIK3CA have been identified innumerous cancers including colorectal, breast, glioblastomas ovarian andlung. In contrast to PIK3CA, no somatic mutations in the β isoform havebeen identified. However, in overexpression studies, the PI3Kβ isoformhas been implicated as necessary for transformation induced by the lossor inactivation of the PTEN tumor suppressor both in vitro and in vivo(Torbett N E, Luna A, Knight Z A, et al., A chemical screen in diversebreast cancer cell lines reveals genetic enhancers and suppressors ofsensitivity to PI3K isotype-selective inhibition. Biochem J2008;415:97-110; Zhao J J, Liu Z, Wang L, Shin E, Loda M F, Roberts T M, Theoncogenic properties of mutant p110a and p110b phosphatidylinositol3-kinases in human mammary epithelial cells. Proc Natl Acad Sci USA2005; 102:18443-8). Consistent with this finding, overexpression of thePIK3CB gene has been identified in some bladder, colon, glioblastomasand leukemias and siRNA mediated knockdown of p110β in glioblastoma celllines results in suppression of tumor growth in vitro and in vivo (Pu P,Kang C, Zhang Z, et al., Downregulation of PIK3CB by siRNA suppressesmalignant glioma cell growth in vitro and in vivo. Technolo Cancer ResTreat 2006; 5:271-280). More recent data using shRNA demonstrated thatdownregulation of p110β and not p110α resulted in PI3K pathwayinactivation and subsequent inactivation of tumor cell growth in PTENdeficient cancers cells both in vitro and in vivo (Wee S, Wiederschain,Maira S-M, Loo A, Miller C, et al., PTEN-deficient cancers depend onPIK3CB. Proc Natl Acad Sci 2008; 105:13057-13062). Consistent with arole of PIK3CB signaling in PTEN null tumors, p110β was reported to beessential to the transformed phenotype in a PTEN-null prostate cancermodel (Jia S, Liu Z, Zhang S, Liu P, Zhang L, et al., Essential roles ofPI(3)K-p110b in cell growth, metabolism and tumorgenesis. Nature 2008;10:1038).

Further, it has been reported that fibrogenesis, including systemicsclerosis (SSc), arthritis, nephropahty, liver cirrhosis, and somecancers, are related to PTEN deficiency and corresponding PI3K-Aktoverexpression (Parapuram, S. K., et al., Loss of PTEN expression bydermal fibroblasts causes skin fibrosis. J. of InvestigativeDermatology, advance online publication 9 Jun. 2011; doi:10.1038/jid.2011.156). Taken together, these findings indicate PI3Kp110β as a promising target for cancer and other syndromes related toPTEN loss (Hollander, M. Christine; Blumenthal, Gideon M.; Dennis,Phillip P.; PTEN loss in the continuum of common cancers, rare syndromesand mouse models. Nature Reviews/Cancer 2011; 11: 289-301). It istherefore desirable to create a potent, selective inhibitor of PI3K-β.

SUMMARY OF THE INVENTION

This invention relates to novel compounds of formula (I):

wherein

-   R1 is selected from H, C₁₋₆alkyl, alkoxy, hydroxy, halogen, —CN,    —NH₂, —NHC(O)Ra, —NHSO₂Ra, —CO₂H, —CO₂Ra, —CONHRb, —CONH₂, —CH₂OH,    and heteroaryl wherein the heteroaryl may be substituted by one or    two C₁₋₃alkyl groups;-   R2 is selected from H, —NHRa, alkoxy, halogen, —CF₃, —CHF₂, and    C₁₋₆alkyl;-   R3 is selected from aryl and heteroaryl, wherein said aryl or    heteroaryl may be substituted by one to three Rc;-   R4 is selected from H or Ra;-   each R5 is independently selected from C₁₋₆alkyl;-   each Ra is independently selected from C₁₋₃alkyl;-   Rb is selected from C₁₋₃alkyl, and SO₂Me;-   each Rc is independently selected from C₁₋₃alkyl, halogen, —CF₃, and    hydroxy; and-   n is 0-2,-   or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention, there is provided a methodof treating a susceptible neoplasm in a mammal in need thereof,comprising administering to the mammal a therapeutically effectiveamount of a compound of formula (I) (including any particularsub-generic formula described herein) or a pharmaceutically acceptablesalt thereof.

In another aspect of the present invention, there is provided a compoundof formula (I), (including any particular sub-generic formula describedherein) or a pharmaceutically acceptable salt thereof for use intherapy.

In another aspect, there is provided a compound of formula (I)(including any particular sub-generic formula described herein) or apharmaceutically acceptable salt thereof for use in the treatment of asusceptible neoplasm in a mammal in need thereof.

In a another aspect of the present invention, there is provided the useof a compound of formula (I) (including any particular sub-genericformula described herein) or a pharmaceutically acceptable salt thereof,in the preparation of a medicament for use in the treatment of asusceptible neoplasm in a mammal in need thereof.

In another aspect of the present invention, there is provided apharmaceutical composition comprising a compound of formula (I)(including any particular sub-generic formula described herein) or apharmaceutically acceptable salt thereof for use in the treatment of asusceptible neoplasm in a mammal in need thereof.

DETAILED DESCRIPTION OF THE INVENTION

This invention is directed to compounds of Formula (I).

According to another embodiment, the invention includes compounds ofFormula (I)(A),

wherein

-   R1 is selected from H, C₁₋₆alkyl, alkoxy, hydroxy, halogen, —CN,    —NH₂, —NHC(O)Ra, —NHSO₂Ra, —CO₂H, —CO₂Ra, —CONHRb, —CONH₂, —CH₂OH,    and heteroaryl wherein the heteroaryl may be substituted by one or    two C₁₋₃alkyl groups, wherein the heteroaryl is selected from the    group consisting of: pyrazolyl, triazolyl, tetrazolyl, oxazolyl and    imidazolyl;-   R2 is selected from H, —NHRa, alkoxy, halogen, —CF₃, —CHF₂, and    C₁₋₆alkyl;-   R3 is selected from aryl and heteroaryl, wherein said aryl or    heteroaryl may be substituted by one to three Rc;-   R4 is selected from H or Ra;-   each R5 is independently selected from C₁₋₆alkyl;-   each Ra is independently selected from C₁₋₃alkyl;-   Rb is selected from C₁₋₃alkyl, and —SO₂Me;-   each Rc is independently selected from C₁₋₃alkyl, halogen, —CF₃, and    hydroxy; and-   n is 0-2,-   or a pharmaceutically acceptable salt thereof.

According to another embodiment, the invention includes compounds ofFormula (I)(B),

wherein

-   R1 is selected from H, C₁₋₆alkyl, alkoxy, hydroxy, halogen, —CN,    —NH2, —NHC(O)Ra, —NHSO₂Ra, —CO₂H, —CO₂Ra, —CONHRb, —CONH₂, —CH₂OH,    and heteroaryl wherein the heteroaryl may be substituted by one or    two C₁₋₃alkyl groups;-   R2 is selected from H, —NHRa, alkoxy, halogen, —CF₃, —CHF₂, and    C₁₋₆alkyl;-   R3 is selected from aryl and heteroaryl, wherein said aryl or    heteroaryl may be substituted by one to three Rc, and wherein the    aryl or heteroaryl are selected from phenyl, naphthyl, benzothienyl,    quinolinyl, isoquinolinyl, and quinazolinyl;-   R4 is selected from H or Ra;-   each R5 is independently selected from C₁₋₆alkyl;-   each Ra is independently selected from C₁₋₃alkyl;-   Rb is selected from C₁₋₃alkyl, and —SO₂Me;-   each Rc is independently selected from C₁₋₃alkyl, halogen, —CF₃, and    hydroxy; and-   n is 0-2,-   or a pharmaceutically acceptable salt thereof.

According to another embodiment, the invention includes compounds ofFormula (I)(B) wherein each Rc is independently C₁₋₃alkyl, F or Cl, andn is 0.

According to another embodiment, the invention includes compounds ofFormula (I)(B) wherein each Rc is independently CF₃ or F, and n is 0.

According to another embodiment, the invention includes the compounds ofFormula (I)(C)

wherein

-   R1 is selected from H, C₁₋₆alkyl, alkoxy, hydroxy, halogen, —CN,    —NHC(O)Ra, —NHSO₂Ra, —CO₂H, —CO₂Ra, —CONHRb, —CONH₂, —CH₂OH, and    heteroaryl wherein the heteroaryl may be substituted by one or two    C₁₋₃alkyl groups;-   R2 is selected from H, —NHRa, alkoxy or C₁₋₆alkyl;-   each of R6, R7, and R8 is independently selected from C₁₋₃alkyl,    halogen, —CF₃, and hydroxyl, or R6 and R7 combine to form a    bi-cyclic aryl or heteroaryl, or R7 and R8 combine to form a    bi-cyclic aryl or heteroaryl;-   each Ra is independently selected from C₁₋₃alkyl; and-   Rb is selected from C₁₋₃alkyl or —SO₂Me;-   or a pharmaceutically acceptable salt thereof.

According to another embodiment, the invention includes the compounds ofFormula (I)(D)

wherein

-   R1 is selected from H, C₁₋₆alkyl, alkoxy, hydroxy, halogen, —CN,    —NHC(O)Ra, —NHSO₂Ra, —CO₂H, —CO₂Ra, —CONHRb, —CONH₂, —CH₂OH, and    heteroaryl wherein the heteroaryl may be substituted by one or two    C₁₋₃alkyl groups;-   R2 is selected from H, NHRa, alkoxy or C₁₋₆alkyl;-   each Ra is independently selected from C₁₋₃alkyl; and-   Rb is selected from C₁₋₃alkyl or SO₂Me;-   or a pharmaceutically acceptable salt thereof.

According to another embodiment, the invention includes the compounds ofFormula (I)(E)

wherein

-   R1 is selected from H, C₁₋₆alkyl, alkoxy, hydroxy, halogen, —CN,    —NHC(O)Ra, —NHSO₂Ra, —CO₂H, —CO₂Ra, —CONHRb, —CONH₂, —CH₂OH, and    heteroaryl wherein the heteroaryl may be substituted by one or two    C₁₋₃alkyl groups;-   R2 is H, NHRa, alkoxy or C₁₋₆alkyl;-   each of R6 and R7 is independently selected from C₁₋₃alkyl, halogen,    —CF₃, and hydroxyl;-   each Ra is independently C₁₋₃alkyl; and-   Rb is C₁₋₃alkyl or SO₂Me;-   or a pharmaceutically acceptable salt thereof.

According to another embodiment, the invention includes compounds:

-   2-(1-methylethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-ol,-   2-ethyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-ol,-   1-[2,3-dichlorophenyl)methyl]-2-(1-methylethyl)-6-(4-morpholinyl)-1H-benzimidazol-4-ol,-   1-[2,3-dichlorophenyl)methyl]-4-fluoro-2-methyl-6-(4-morpholinyl)-1H-benzimidazole,-   1-[2,3-dichlorophenyl)methyl]-2-ethyl-6-(4-morpholinyl)-1H-benzimidazol-4-ol,-   4-fluoro-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole,-   2-ethyl-4-fluoro-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole,-   2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[2,3-dichlorophenyl)methyl]-2-ethyl-4-fluoro-6-(4-morpholinyl)-1H-benzimidazole,-   2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-pyrazol-5-yl)-1H-benzimidazole,-   1-[2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-4-(1H-pyrazol-5-yl)-1H-benzimidazole,-   2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazole,-   methyl    2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylate,-   1-[2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide,-   methyl    1-[(2-fluoro-3-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,-   2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carbonitrile,-   1-[2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carbonitrile,-   methyl    2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,-   2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[(2-fluoro-3-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide,-   1-[2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazole,-   methyl    2-methyl-6-(4-morpholinyl)-1-(5-quinolinylmethyl)-1H-benzimidazole-4-carboxylate,-   2-methyl-6-(4-morpholinyl)-1-(5-quinolinylmethyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[3,4-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   2-methyl-6-(4-morpholinyl)-1-(2-naphthalenylmethyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[(3,4-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazole,-   2-methyl-4-(3-methyl-1H-1,2,4-triazol-5-yl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole,-   1-[2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-yl]ethanone,-   [2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-yl]methanol,-   2-methyl-N-(methylsulfonyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide,-   methyl    5-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-7-carboxylate,-   methyl    1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate,-   1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylic    acid,-   6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylic    acid,-   methyl    1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate,-   1-[2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylic    acid,-   1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxamide,-   methyl    6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate,-   methyl    1-[(2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate,-   1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-2-(trifluoromethyl)-1H-benzimidazole,-   1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-2-(trifluoromethyl)-1H-benzimidazole,-   1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-2-(trifluoromethyl)-1H-benzimidazole,-   2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-tetrazol-5-yl)-1H-benzimidazole,-   [2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazol-4-yl]methanol,-   1-[(3-chloro-2-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   2-methyl-1-[(2-methylphenyl)methyl]-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   ethyl    2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,-   4-bromo-2-methyl-6-(4-morpholinyl)-1H-benzimidazole,-   4-bromo-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole,-   2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1,3-oxazol-2-yl)-1H-benzimidazole,-   methyl 2-chloro-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate,-   methyl    2-chloro-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,-   2-chloro-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid-   methyl    2-(difluoromethyl)-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate,-   2-(difluoromethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylic    acid,-   2-(difluoromethyl)-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[2,3-dichlorophenyl)methyl]-2-(difluoromethyl)-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[3-chloro-2-methylphenyl)methyl]-2-(difluoromethyl)-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   1-(1-benzothien-7-ylmethyl)-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[2,3-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   1-[3-fluoro-2-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   2,4-dimethyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole,-   1-[1-(3-chloro-2-methylphenyl)ethyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylic    acid,-   2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1,3-thiazol-2-yl)-1H-benzimidazole,-   4-(2-furanyl)-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole    and-   2-methyl-4-[(methyloxy)methyl]-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole.

DEFINITIONS

By the term “aryl” as used herein, unless otherwise defined, is meantaromatic, hydrocarbon, ring system. The ring system may be monocyclic orfused polycyclic (e.g. bicyclic, tricyclic, etc.). In variousembodiments, the monocyclic aryl ring is C5-C10, or C5-C7, or C5-C6,where these carbon numbers refer to the number of carbon atoms that formthe ring system. A C6 ring system, i.e. a phenyl ring is a suitable arylgroup. In various embodiments, the polycyclic ring is a bicyclic arylgroup, where suitable bicyclic aryl groups are C8-C12, or C9-C10. Anaphthyl ring, which has 10 carbon atoms, is a suitable polycyclic arylgroup.

By the term “heteroaryl” as used herein, unless otherwise defined, ismeant an aromatic ring system containing carbon(s) and at least oneheteroatom. Heteroaryl may be monocyclic or polycyclic. A monocyclicheteroaryl group may have 1 to 4 heteroatoms in the ring, while apolycyclic heteroaryl may contain 1 to 10 hetero atoms. A polycyclicheteroaryl ring may contain fused, spiro or bridged ring junctions, forexample, bicyclic heteroaryl is a polycyclic heteroaryl. Bicyclicheteroaryl rings may contain from 8 to 12 member atoms. Monocyclicheteroaryl rings may contain from 5 to 8 member atoms (carbons andheteroatoms). Exemplary heteroaryl groups include: benzofuran,benzothiene, benzothiophene, furan, imidazole, indole, isothiazole,oxazole, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole,quinoline, isoquinoline, quinazoline, quinoxaline, thiazole, andthiophene. According to an alternative embodiment, heteroaryls may besubstituted with one to three alkyl groups.

By the term “alkoxy” as used herein is meant —O(alkyl) including —OCH₃,—OCH₂CH₃ and —OC(CH₃)₃ where alkyl is as described herein.

By the term “heteroatom” as used herein is meant oxygen, nitrogen orsulfur.

By the term “halogen” as used herein is meant a substituent selectedfrom bromide, iodide, chloride and fluoride.

By the term “alkyl” and derivatives thereof and in all carbon chains asused herein, including alkyl chains defined by the term “—(CH₂)_(n)”,“—(CH₂)_(m)” and the like, is meant a linear or branched, saturated orunsaturated hydrocarbon chain, and unless otherwise defined, the carbonchain will contain from 1 to 12 carbon atoms.

By the term “co-administering” and derivatives thereof as used herein ismeant either simultaneous administration or any manner of separatesequential administration of a PI3 kinase inhibiting compound, asdescribed herein, and a further active ingredient or ingredients. Theterm further active ingredient or ingredients, as used herein, includesany compound or therapeutic agent known to or that demonstratesadvantageous properties when administered to a patient in need oftreatment. Suitably, if the administration is not simultaneous, thecompounds are administered in a close time proximity to each other.Furthermore, it does not matter if the compounds are administered in thesame dosage form, e.g. one compound may be administered topically andanother compound may be administered orally.

The term “compound” as used herein includes all isomers of the compound.Examples of such isomers include: enantiomers, tautomers, rotamers.

In formulas where a “dotted” bond is drawn between two atoms, it ismeant that such bond can be either single or double bond. A ring systemcontaining such bonds can be aromatic or non-aromatic.

Certain compounds described herein may contain one or more chiral atoms,or may otherwise be capable of existing as two enantiomers, or two ormore diastereoisomers. Accordingly, the compounds of this inventioninclude mixtures of enantiomers/diastereoisomers as well as purifiedenantiomers/diastereoisomers or enantiomerically/diastereoisomericallyenriched mixtures. Also included within the scope of the invention arethe individual isomers of the compounds represented by Formula (I) aboveas well as any wholly or partially equilibrated mixtures thereof. Thepresent invention also covers the individual isomers of the compoundsrepresented by the formulas above as mixtures with isomers thereof inwhich one or more chiral centers are inverted. The present inventionalso includes isotopomers of the compounds of Formula (I). Examples ofsuch isotopomers include but not limited to compounds with one of moredeuterium atoms.

Compounds of Formula (I) are included in the pharmaceutical compositionsof the invention. Where a —COOH or —OH group is present,pharmaceutically acceptable esters can be employed, for example methyl,ethyl, pivaloyloxymethyl, and the like for —COOH, and acetate maleateand the like for —OH, and those esters known in the art for modifyingsolubility or hydrolysis characteristics, for use as sustained releaseor prodrug formulations.

It will be appreciated by those skilled in the art that the compounds offormula (I) may be utilized as a pharmaceutically acceptable saltversion thereof. The pharmaceutically acceptable salts of the compoundsof formula (I) include conventional salts formed from pharmaceuticallyacceptable (i.e., non-toxic) inorganic or organic acids or bases as wellas quaternary ammonium salts. Representative salts include thefollowing: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate,bitartrate, borate, bromide, calcium edetate, camsylate, carbonate,chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate,estolate, esylate, ethanol amine, fumarate, gluceptate, gluconate,glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine,hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate,lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate(methanesulfonate), methylbromide, methylnitrate, methylsulfate,monopotassium maleate, mucate, napsylate, nitrate, N-methylglucamine,oxalate, pamoate (embonate), palmitate, pantothenate,phosphate/diphosphate, polygalacturonate, potassium, salicylate, sodium,stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate(methylbenzenesulfonate), triethiodide, trimethylammonium and valerate.Other salts, such as oxalic and trifluoroacetic, which are notthemselves pharmaceutically acceptable, may be useful in the preparationof salts useful as intermediates in obtaining compounds of thisinvention and these form a further aspect of the invention. In oneembodiment, the compound of formula (I) is in the form of the free base.In one embodiment, the compound of formula (I) is in the form of thetris salt, i.e. tris(hydroxymethyl)aminomethane. In one embodiment, thecompound of formula (I) is in the form of the sulfate salt. In oneembodiment, the compound of formula (I) is in the form of thehydrochloride salt. In one embodiment, the compound of formula (I) is inthe form of the sodium salt. Certain salt versions of the compounds maybe solvates, particularly hydrates. In one embodiment, the compound offormula (I) or a pharmaceutically acceptable salt thereof is in the formof a mono-, di-, tri- or hemi-hydrate.

It has now been found that compounds of the present invention areinhibitors of the Phosphatoinositides 3-kinases (PI3Ks). When thephosphatoinositides 3-kinase (PI3K) enzyme is inhibited by a compound ofthe present invention, PI3K is unable to exert its enzymatic, biologicaland/or pharmacological effects. The compounds of the present inventionare therefore useful in the treatment of autoimmune disorders,inflammatory diseases, cardiovascular diseases, neurodegenerativediseases, allergy, asthma, pancreatitis, multiorgan failure, kidneydiseases, platelet aggregation, cancer, sperm motility, transplantationrejection, graft rejection and lung injuries.

Compounds according to Formula (I) are suitable for the modulation,notably the inhibition of the activity of phosphatoinositides 3-kinases(PI3K) and, more particularly, selective inhibitors of the beta isoformof phosphatoinositides 3-kinase (PI3Kβ). Therefore the compounds of thepresent invention are also useful for the treatment of disorders whichare mediated by PI3Ks. Said treatment involves the modulation—notablythe inhibition or the down regulation—of the phosphatoinositides3-kinases.

Because the pharmaceutically active compounds of the present inventionare active as PI3 kinase inhibitors, particularly the compounds thatinhibit PI3Kβ, either selectively or in conjunction with one or more ofPI3δ, PI3Kα, and/or PI3Kγ, they exhibit therapeutic utility in treatmentof susceptible neoplasms, particularly those neoplasms that exhibit aPTEN deficiency.

As used herein, the phrase “PTEN deficient” or “PTEN deficiency” shalldescribe tumors with deficiencies of the tumor suppressor function ofPTEN (Phosphatase and Tensin Homolog). Such deficiency includes mutationin the PTEN gene, reduction or absence of PTEN proteins when compared toPTEN wild-type, or mutation or absence of other genes that causesuppression of PTEN function.

As used herein, the term “treatment” or “treating” in the context oftherapeutic methods, refers to alleviating the specified condition,eliminating or reducing the symptoms of the condition, slowing oreliminating the progression, invasion, or metastatic spread of thecondition and preventing or delaying the reoccurrence of the conditionin a previously afflicted subject. The present invention furtherprovides use of the compounds of the invention for the preparation of amedicament for the treatment of several conditions in a mammal (e.g.,human) in need thereof.

“Susceptible neoplasm” as used herein refers to neoplasms which aresusceptible to treatment by a kinase inhibitor and particularlyneoplasms that are susceptible to treatment by a PI3Kβ inhibitor.Neoplasms which have been associated with inappropriate activity of thePTEN phosphatase and particularly neoplasms which are exhibit mutationof PTEN, or mutation of an upstream activator of PI3Kβ kinase oroverexpression of an upstream activator of PI3Kβ kinase, and aretherefore susceptible to treatment with an PI3Kβ inhibitor are known inthe art, and include both primary and metastatic tumors and cancers.According to one embodiment, description of the treatment of asusceptible neoplasm may be used interchangeably with description of thetreatment of a cancer.

According to one embodiment, “susceptible neoplasms” includes, but arenot limited to PTEN-deficient neoplasms listed as follows:

brain (gliomas),

glioblastomas,

leukemias,

Bannayan-Zonana syndrome,

Cowden disease,

Lhermitte-Duclos disease,

breast cancer,

inflammatory breast cancer,

colorectal cancer

Wilm's tumor,

Ewing's sarcoma,

Rhabdomyosarcoma,

ependymoma,

medulloblastoma,

colon cancer,

head and neck cancer,

kidney cancer,

lung cancer,

liver cancer,

melanoma,

squamous cell carcinoma,

ovarian cancer,

pancreatic cancer,

prostate cancer,

sarcoma cancer,

osteosarcoma,

giant cell tumor of bone,

thyroid cancer,

lymphoblastic T cell leukemia,

chronic myelogenous leukemia,

chronic lymphocytic leukemia,

hairy-cell leukemia,

acute lymphoblastic leukemia,

acute myelogenous leukemia,

chronic neutrophilic leukemia,

acute lymphoblastic T cell leukemia,

Plasmacytoma,

Immunoblastic large cell leukemia,

Mantle cell leukemia,

Multiple myeloma,

Megakaryoblastic leukemia,

multiple myeloma,

Acute megakaryocytic leukemia,

promyelocytic leukemia,

Erythroleukemia,

malignant lymphoma,

hodgkins lymphoma,

non-hodgkins lymphoma,

lymphoblastic T cell lymphoma,

Burkitt's lymphoma,

follicular lymphoma,

neuroblastoma,

bladder cancer,

urothelial cancer,

vulval cancer,

cervical cancer,

endometrial cancer,

renal cancer,

mesothelioma,

esophageal cancer,

salivary gland cancer,

hepatocellular cancer,

gastric cancer,

nasopharangeal cancer,

buccal cancer,

cancer of the mouth,

GIST (gastrointestinal stromal tumor),

and testicular cancer.

According to an alternative embodiment, the term “susceptible neoplasm”includes and is limited to hormone refractory prostate cancer,non-small-cell lung cancer, endometrial cancer, gastric cancer,melanoma, head and neck cancer, breast cancer, including trip-negativebreast cancer, and glioma. PTEN deficiency has been correlated to suchcancers as demonstrated in a number of published resources, e.g. Am JClin Pathol. 2009 February; 131(2):257-63 (glioblastoma), J ClinNeurosci. 2010 December; 17(12): 1543-7 (glioblastoma), Nat. Genet. 2009May; 41(5):619-24 (prostate cancer), Br J Cancer. 2008 Oct. 21;99(8):1296-301 (prostate cancer), Int J Cancer. 2007 Mar. 15;120(6):1284-92 (prostate cancer), J Invest Dermatol. 2006 January;126(1):154-60 (melanoma), J Clin Oncol. 2006 Jan. 10; 24(2):288-95(melanoma), Am J Clin Pathol. 2005 October; 124(4):528-36 (melanoma),Int J Oncol. 2009 April; 34(4):983-93 (breast cancer), Epigenetics. 2011May 1; 6(5):638-49 (breast cancer), Gynecol Oncol. 2009 February;112(2):307-13 (ovarian cancer), Mod Pathol. 2010 October; 23(10):1316-24(ovarian cancer), J Pathol. 2010 February; 220(3):392-400 (ovariancancer), Lung. 2009 March-April; 187(2):104-9 (lung cancer), AnticancerRes. 2007 January-February; 27(1B):575-81 (lung cancer), Am J. Surg.2008 June; 195(6):719-25 (colon cancer), J Clin Oncol. 2009 Dec. 10;27(35):5924-30 (colon cancer), Gynecol Oncol. 2004 June; 93(3):621-7(cervical cancer), and J Oral Pathol Med. 2002 August; 31(7):379-84(head and neck cancer).

In another aspect of the present invention, there is provided a methodof treating a susceptible neoplasm in a mammal in need thereof,comprising administering to the mammal a therapeutically effectiveamount of a compound of formula (I) (including any particularsub-generic formula described herein) or a pharmaceutically acceptablesalt thereof.

In another aspect of the present invention, there is provided a methodof treating fibrosis in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of acompound of formula (I) (including any particular sub-generic formuladescribed herein) or a pharmaceutically acceptable salt thereof.Fibrosis includes, alternatively or collectively, systemic sclerosis(SSc), arthritis, nephropahty, and liver cirrhosis.

In another aspect of the present invention, there is provided a methodof treating hormone refractory prostate cancer in a mammal in needthereof, comprising administering to the mammal a therapeuticallyeffective amount of a compound of formula (I) (including any particularsub-generic formula described herein) or a pharmaceutically acceptablesalt thereof.

In another aspect of the present invention, there is provided a methodof treating non-small-cell lung cancer in a mammal in need thereof,comprising administering to the mammal a therapeutically effectiveamount of a compound of formula (I) (including any particularsub-generic formula described herein) or a pharmaceutically acceptablesalt thereof.

In another aspect of the present invention, there is provided a methodof treating endometrial cancer in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of acompound of formula (I) (including any particular sub-generic formuladescribed herein) or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention, there is provided a methodof treating gastric cancer in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of acompound of formula (I) (including any particular sub-generic formuladescribed herein) or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention, there is provided a methodof treating melanoma in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of acompound of formula (I) (including any particular sub-generic formuladescribed herein) or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention, there is provided a methodof treating head and neck cancer in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of acompound of formula (I) (including any particular sub-generic formuladescribed herein) or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention, there is provided a methodof treating trip-negative breast cancer in a mammal in need thereof,comprising administering to the mammal a therapeutically effectiveamount of a compound of formula (I) (including any particularsub-generic formula described herein) or a pharmaceutically acceptablesalt thereof.

In another aspect of the present invention, there is provided a methodof treating glioma in a mammal in need thereof, comprising administeringto the mammal a therapeutically effective amount of a compound offormula (I) (including any particular sub-generic formula describedherein) or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention, there is provided a compoundof formula (I), (including any particular sub-generic formula describedherein) or a pharmaceutically acceptable salt thereof for use intherapy.

In another aspect, there is provided a compound of formula (I)(including any particular sub-generic formula described herein) or apharmaceutically acceptable salt thereof for use in the treatment of asusceptible neoplasm in a mammal in need thereof.

In a another aspect of the present invention, there is provided the useof a compound of formula (I) (including any particular sub-genericformula described herein) or a pharmaceutically acceptable salt thereof,in the preparation of a medicament for use in the treatment of asusceptible neoplasm in a mammal in need thereof.

In another aspect of the present invention, there is provided apharmaceutical composition comprising a compound of formula (I)(including any particular sub-generic formula described herein) or apharmaceutically acceptable salt thereof for use in the treatment of asusceptible neoplasm in a mammal in need thereof.

When a compound of Formula (I) is administered for the treatment ofcancer, the term “co-administering” and derivatives thereof as usedherein is meant either simultaneous administration or any manner ofseparate sequential administration of a PI3 kinase inhibiting compound,as described herein, and a further active ingredient or ingredients,known to be useful in the treatment of cancer, including chemotherapyand radiation treatment. The term further active ingredient oringredients, as used herein, includes any compound or therapeutic agentknown to or that demonstrates advantageous properties when administeredto a patient in need of treatment for cancer. Preferably, if theadministration is not simultaneous, the compounds are administered in aclose time proximity to each other. Furthermore, it does not matter ifthe compounds are administered in the same dosage form, e.g. onecompound may be administered topically and another compound may beadministered orally.

Typically, any anti-neoplastic agent that has activity versus asusceptible tumor being treated may be co-administered in the treatmentof cancer in the present invention. Examples of such agents can be foundin Cancer Principles and Practice f Oncology by V. T. Devita and S.Hellman (editors), 6^(th) edition (Feb. 15, 2001), Lippincott Williams &Wilkins Publishers. A person of ordinary skill in the art would be ableto discern which combinations of agents would be useful based on theparticular characteristics of the drugs and the cancer involved. Typicalanti-neoplastic agents useful in the present invention include, but arenot limited to, anti-microtubule agents such as diterpenoids and vincaalkaloids; platinum coordination complexes; alkylating agents such asnitrogen mustards, oxazaphosphorines, alkylsulfonates, nitrosoureas, andtriazenes; antibiotic agents such as anthracyclins, actinomycins andbleomycins; topoisomerase II inhibitors such as epipodophyllotoxins;antimetabolites such as purine and pyrimidine analogues and anti-folatecompounds; topoisomerase I inhibitors such as camptothecins; hormonesand hormonal analogues; signal transduction pathway inhibitors;non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeuticagents; proapoptotic agents; and cell cycle signaling inhibitors.

Examples of a further active ingredient or ingredients for use incombination or co-administered with the present PI3 kinase inhibitingcompounds are chemotherapeutic agents.

Anti-microtubule or anti-mitotic agents are phase specific agents activeagainst the microtubules of tumor cells during M or the mitosis phase ofthe cell cycle. Examples of anti-microtubule agents include, but are notlimited to, diterpenoids and vinca alkaloids.

Diterpenoids, which are derived from natural sources, are phase specificanti-cancer agents that operate at the G₂/M phases of the cell cycle. Itis believed that the diterpenoids stabilize the β-tubulin subunit of themicrotubules, by binding with this protein. Disassembly of the proteinappears then to be inhibited with mitosis being arrested and cell deathfollowing. Examples of diterpenoids include, but are not limited to,paclitaxel and its analog docetaxel.

Paclitaxel, 5β,20-epoxy-1,2α,4,7β,10β,13α-hexa-hydroxytax-11-en-9-one4,10-diacetate 2-benzoate 13-ester with(2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene productisolated from the Pacific yew tree Taxus brevifolia and is commerciallyavailable as an injectable solution TAXOL®. It is a member of the taxanefamily of terpenes. It was first isolated in 1971 by Wani et al. J. Am.Chem., Soc., 93:2325. 1971), who characterized its structure by chemicaland X-ray crystallographic methods. One mechanism for its activityrelates to paclitaxel's capacity to bind tubulin, thereby inhibitingcancer cell growth. Schiff et al., Proc. Natl. Acad, Sci. USA,77:1561-1565 (1980); Schiff et al., Nature, 277:665-667 (1979); Kumar,J. Biol, Chem, 256: 10435-10441 (1981). For a review of synthesis andanticancer activity of some paclitaxel derivatives see: D. G. I.Kingston et al., Studies in Organic Chemistry vol. 26, entitled “Newtrends in Natural Products Chemistry 1986”, Attaur-Rahman, P. W. LeQuesne, Eds. (Elsevier, Amsterdam, 1986) pp 219-235.

Paclitaxel has been approved for clinical use in the treatment ofrefractory ovarian cancer in the United States (Markman et al., YaleJournal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann.Intem, Med., 111:273, 1989) and for the treatment of breast cancer(Holmes et al., J. Nat. Cancer Inst., 83:1797, 1991.) It is a potentialcandidate for treatment of neoplasms in the skin (Einzig et. al., Proc.Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastireet. al., Sem. Oncol., 20:56, 1990). The compound also shows potentialfor the treatment of polycystic kidney disease (Woo et. al., Nature,368:750. 1994), lung cancer and malaria. Treatment of patients withpaclitaxel results in bone marrow suppression (multiple cell lineages,Ignoff, R. J. et. al, Cancer Chemotherapy Pocket Guide, 1998) related tothe duration of dosing above a threshold concentration (50 nM) (Kearns,C. M. et. al., Seminars in Oncology, 3(6) p. 16-23, 1995).

Docetaxel, (2R,3S)-N-carboxy-3-phenylisoserine,N-tert-butyl ester,13-ester with 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one4-acetate 2-benzoate, trihydrate; is commercially available as aninjectable solution as TAXOTERE®. Docetaxel is indicated for thetreatment of breast cancer. Docetaxel is a semisynthetic derivative ofpaclitaxel q.v., prepared using a natural precursor,10-deacetyl-baccatin III, extracted from the needle of the European Yewtree. The dose limiting toxicity of docetaxel is neutropenia.

Vinca alkaloids are phase specific anti-neoplastic agents derived fromthe periwinkle plant. Vinca alkaloids act at the M phase (mitosis) ofthe cell cycle by binding specifically to tubulin. Consequently, thebound tubulin molecule is unable to polymerize into microtubules.Mitosis is believed to be arrested in metaphase with cell deathfollowing. Examples of vinca alkaloids include, but are not limited to,vinblastine, vincristine, and vinorelbine.

Vinblastine, vincaleukoblastine sulfate, is commercially available asVELBAN® as an injectable solution. Although, it has possible indicationas a second line therapy of various solid tumors, it is primarilyindicated in the treatment of testicular cancer and various lymphomasincluding Hodgkin's Disease; and lymphocytic and histiocytic lymphomas.Myelosuppression is the dose limiting side effect of vinblastine.

Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commerciallyavailable as ONCOVIN® as an injectable solution. Vincristine isindicated for the treatment of acute leukemias and has also found use intreatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas.Alopecia and neurologic effects are the most common side effect ofvincristine and to a lesser extent myelosupression and gastrointestinalmucositis effects occur.

Vinorelbine,3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine[R—(R*,R*)-2,3-dihydroxybutanedioate(1:2)(salt)], commercially available as an injectable solution ofvinorelbine tartrate (NAVELBINE®), is a semisynthetic vinca alkaloid.Vinorelbine is indicated as a single agent or in combination with otherchemotherapeutic agents, such as cisplatin, in the treatment of varioussolid tumors, particularly non-small cell lung, advanced breast, andhormone refractory prostate cancers. Myelosuppression is the most commondose limiting side effect of vinorelbine.

Platinum coordination complexes are non-phase specific anti-canceragents, which are interactive with DNA. The platinum complexes entertumor cells, undergo, aquation and form intra- and interstrandcrosslinks with DNA causing adverse biological effects to the tumor.Examples of platinum coordination complexes include, but are not limitedto, cisplatin and carboplatin.

Cisplatin, cis-diamminedichloroplatinum, is commercially available asPLATINOL® as an injectable solution. Cisplatin is primarily indicated inthe treatment of metastatic testicular and ovarian cancer and advancedbladder cancer. The primary dose limiting side effects of cisplatin arenephrotoxicity, which may be controlled by hydration and diuresis, andototoxicity.

Carboplatin, platinum, diammine[1,1-cyclobutane-dicarboxylate(2-)-O,O′],is commercially available as PARAPLATIN® as an injectable solution.Carboplatin is primarily indicated in the first and second linetreatment of advanced ovarian carcinoma. Bone marrow suppression is thedose limiting toxicity of carboplatin.

Alkylating agents are non-phase anti-cancer specific agents and strongelectrophiles. Typically, alkylating agents form covalent linkages, byalkylation, to DNA through nucleophilic moieties of the DNA moleculesuch as phosphate, amino, sulfhydryl, hydroxy, carboxyl, and imidazolegroups. Such alkylation disrupts nucleic acid function leading to celldeath. Examples of alkylating agents include, but are not limited to,nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil;alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; andtriazenes such as dacarbazine.

Cyclophosphamide,2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxidemonohydrate, is commercially available as an injectable solution ortablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent orin combination with other chemotherapeutic agents, in the treatment ofmalignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea,vomiting and leukopenia are the most common dose limiting side effectsof cyclophosphamide.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commerciallyavailable as an injectable solution or tablets as ALKERAN®. Melphalan isindicated for the palliative treatment of multiple myeloma andnon-respectable epithelial carcinoma of the ovary. Bone marrowsuppression is the most common dose limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, iscommercially available as LEUKERAN® tablets. Chlorambucil is indicatedfor the palliative treatment of chronic lymphatic leukemia, andmalignant lymphomas such as lymphosarcoma, giant follicular lymphoma,and Hodgkin's disease. Bone marrow suppression is the most common doselimiting side effect of chlorambucil.

Busulfan, 1,4-butanediol dimethanesulfonate, is commercially availableas MYLERAN® TABLETS. Busulfan is indicated for the palliative treatmentof chronic myelogenous leukemia. Bone marrow suppression is the mostcommon dose limiting side effects of busulfan.

Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is commerciallyavailable as single vials of lyophilized material as BiCNU®. Carmustineis indicated for the palliative treatment as a single agent or incombination with other agents for brain tumors, multiple myeloma,Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppressionis the most common dose limiting side effects of carmustine.

Dacarbazine, 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, iscommercially available as single vials of material as DTIC-Dome®.Dacarbazine is indicated for the treatment of metastatic malignantmelanoma and in combination with other agents for the second linetreatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are themost common dose limiting side effects of dacarbazine.

Antibiotic anti-neoplastics are non-phase specific agents, which bind orintercalate with DNA. Typically, such action results in stable DNAcomplexes or strand breakage, which disrupts ordinary function of thenucleic acids leading to cell death. Examples of antibioticanti-neoplastic agents include, but are not limited to, actinomycinssuch as dactinomycin, anthrocyclins such as daunorubicin anddoxorubicin; and bleomycins.

Dactinomycin, also know as Actinomycin D, is commercially available ininjectable form as COSMEGEN®. Dactinomycin is indicated for thetreatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, andanorexia are the most common dose limiting side effects of dactinomycin.

Daunorubicin,(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as a liposomalinjectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.Daunorubicin is indicated for remission induction in the treatment ofacute nonlymphocytic leukemia and advanced HIV associated Kaposi'ssarcoma. Myelosuppression is the most common dose limiting side effectof daunorubicin.

Doxorubicin,(8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-8-glycoloyl,7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12 naphthacenedionehydrochloride, is commercially available as an injectable form as RUBEX®or ADRIAMYCIN RDF®. Doxorubicin is primarily indicated for the treatmentof acute lymphoblastic leukemia and acute myeloblastic leukemia, but isalso a useful component in the treatment of some solid tumors andlymphomas. Myelosuppression is the most common dose limiting side effectof doxorubicin.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated froma strain of Streptomyces verticillus, is commercially available asBLENOXANE®. Bleomycin is indicated as a palliative treatment, as asingle agent or in combination with other agents, of squamous cellcarcinoma, lymphomas, and testicular carcinomas Pulmonary and cutaneoustoxicities are the most common dose limiting side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to,epipodophyllotoxins.

Epipodophyllotoxins are phase specific anti-neoplastic agents derivedfrom the mandrake plant. Epipodophyllotoxins typically affect cells inthe S and G₂ phases of the cell cycle by forming a ternary complex withtopoisomerase II and DNA causing DNA strand breaks. The strand breaksaccumulate and cell death follows. Examples of epipodophyllotoxinsinclude, but are not limited to, etoposide and teniposide.

Etoposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially availableas an injectable solution or capsules as VePESID® and is commonly knownas VP-16. Etoposide is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of testicular andnon-small cell lung cancers. Myelosuppression is the most common sideeffect of etoposide. The incidence of leucopenia tends to be more severethan thrombocytopenia.

Teniposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-thenylidene-β-D-glucopyranoside], is commercially availableas an injectable solution as VUMON® and is commonly known as VM-26.Teniposide is indicated as a single agent or in combination with otherchemotherapy agents in the treatment of acute leukemia in children.Myelosuppression is the most common dose limiting side effect ofteniposide. Teniposide can induce both leucopenia and thrombocytopenia.

Antimetabolite neoplastic agents are phase specific anti-neoplasticagents that act at S phase (DNA synthesis) of the cell cycle byinhibiting DNA synthesis or by inhibiting purine or pyrimidine basesynthesis and thereby limiting DNA synthesis. Consequently, S phase doesnot proceed and cell death follows. Examples of antimetaboliteanti-neoplastic agents include, but are not limited to, fluorouracil,methotrexate, cytarabine, mercaptopurine, thioguanine, and gemcitabine.

5-fluorouracil, 5-fluoro-2,4-(1H,3H) pyrimidinedione, is commerciallyavailable as fluorouracil. Administration of 5-fluorouracil leads toinhibition of thymidylate synthesis and is also incorporated into bothRNA and DNA. The result typically is cell death. 5-fluorouracil isindicated as a single agent or in combination with other chemotherapyagents in the treatment of carcinomas of the breast, colon, rectum,stomach and pancreas. Myelosuppression and mucositis are dose limitingside effects of 5-fluorouracil. Other fluoropyrimidine analogs include5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridinemonophosphate.

Cytarabine, 4-amino-1-β-D-arabinofuranosyl-2(1H)-pyrimidinone, iscommercially available as CYTOSAR-U® and is commonly known as Ara-C. Itis believed that cytarabine exhibits cell phase specificity at S-phaseby inhibiting DNA chain elongation by terminal incorporation ofcytarabine into the growing DNA chain. Cytarabine is indicated as asingle agent or in combination with other chemotherapy agents in thetreatment of acute leukemia. Other cytidine analogs include5-azacytidine and 2′,2′-difluorodeoxycytidine (gemcitabine). Cytarabineinduces leucopenia, thrombocytopenia, and mucositis.

Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, iscommercially available as PURINETHOL®. Mercaptopurine exhibits cellphase specificity at S-phase by inhibiting DNA synthesis by an as of yetunspecified mechanism. Mercaptopurine is indicated as a single agent orin combination with other chemotherapy agents in the treatment of acuteleukemia. Myelosuppression and gastrointestinal mucositis are expectedside effects of mercaptopurine at high doses. A useful mercaptopurineanalog is azathioprine.

Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is commerciallyavailable as TABLOID®. Thioguanine exhibits cell phase specificity atS-phase by inhibiting DNA synthesis by an as of yet unspecifiedmechanism. Thioguanine is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of acute leukemia.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of thioguanine administration.However, gastrointestinal side effects occur and can be dose limiting.Other purine analogs include pentostatin, erythrohydroxynonyladenine,fludarabine phosphate, and cladribine.

Gemcitabine, 2′-deoxy-2′,2′-difluorocytidine monohydrochloride(β-isomer), is commercially available as GEMZAR®. Gemcitabine exhibitscell phase specificity at S-phase and by blocking progression of cellsthrough the G1/S boundary. Gemcitabine is indicated in combination withcisplatin in the treatment of locally advanced non-small cell lungcancer and alone in the treatment of locally advanced pancreatic cancer.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of gemcitabine administration.

Methotrexate,N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamicacid, is commercially available as methotrexate sodium. Methotrexateexhibits cell phase effects specifically at S-phase by inhibiting DNAsynthesis, repair and/or replication through the inhibition ofdyhydrofolic acid reductase which is required for synthesis of purinenucleotides and thymidylate. Methotrexate is indicated as a single agentor in combination with other chemotherapy agents in the treatment ofchoriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, andcarcinomas of the breast, head, neck, ovary and bladder.Myelosuppression (leucopenia, thrombocytopenia, and anemia) andmucositis are expected side effect of methotrexate administration.

Camptothecins, including, camptothecin and camptothecin derivatives areavailable or under development as Topoisomerase I inhibitors.Camptothecins cytotoxic activity is believed to be related to itsTopoisomerase I inhibitory activity. Examples of camptothecins include,but are not limited to irinotecan, topotecan, and the various opticalforms of7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecindescribed below.

Irinotecan HCl, (4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride, is commercially available as the injectable solutionCAMPTOSAR®.

Irinotecan is a derivative of camptothecin which binds, along with itsactive metabolite SN-38, to the topoisomerase I-DNA complex. It isbelieved that cytotoxicity occurs as a result of irreparable doublestrand breaks caused by interaction of the topoisomerase I:DNA:irintecanor SN-38 ternary complex with replication enzymes. Irinotecan isindicated for treatment of metastatic cancer of the colon or rectum. Thedose limiting side effects of irinotecan HCl are myelosuppression,including neutropenia, and GI effects, including diarrhea.

Topotecan HCl,(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dionemonohydrochloride, is commercially available as the injectable solutionHYCAMTIN®. Topotecan is a derivative of camptothecin which binds to thetopoisomerase I-DNA complex and prevents religation of singles strandbreaks caused by Topoisomerase I in response to torsional strain of theDNA molecule. Topotecan is indicated for second line treatment ofmetastatic carcinoma of the ovary and small cell lung cancer. The doselimiting side effect of topotecan HCl is myelosuppression, primarilyneutropenia.

Also of interest, is the camptothecin derivative of formula A following,currently under development, including the racemic mixture (R,S) form aswell as the R and S enantiomers:

known by the chemical name“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R,S)-camptothecin(racemic mixture) or“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R)-camptothecin(R enantiomer) or“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(S)-camptothecin(S enantiomer). Such compound as well as related compounds aredescribed, including methods of making, in U.S. Pat. Nos. 6,063,923;5,342,947; 5,559,235; 5,491,237 and pending U.S. patent application Ser.No. 08/977,217 filed Nov. 24, 1997.

Hormones and hormonal analogues are useful compounds for treatingcancers in which there is a relationship between the hormone(s) andgrowth and/or lack of growth of the cancer. Examples of hormones andhormonal analogues useful in cancer treatment include, but are notlimited to, adrenocorticosteroids such as prednisone and prednisolonewhich are useful in the treatment of malignant lymphoma and acuteleukemia in children; aminoglutethimide and other aromatase inhibitorssuch as anastrozole, letrazole, vorazole, and exemestane useful in thetreatment of adrenocortical carcinoma and hormone dependent breastcarcinoma containing estrogen receptors; progestrins such as megestrolacetate useful in the treatment of hormone dependent breast cancer andendometrial carcinoma; estrogens, androgens, and anti-androgens such asflutamide, nilutamide, bicalutamide, cyproterone acetate and5α-reductases such as finasteride and dutasteride, useful in thetreatment of prostatic carcinoma and benign prostatic hypertrophy;anti-estrogens such as tamoxifen, toremifene, raloxifene, droloxifene,iodoxyfene, as well as selective estrogen receptor modulators (SERMS)such those described in U.S. Pat. Nos. 5,681,835, 5,877,219, and6,207,716, useful in the treatment of hormone dependent breast carcinomaand other susceptible cancers; and gonadotropin-releasing hormone (GnRH)and analogues thereof which stimulate the release of leutinizing hormone(LH) and/or follicle stimulating hormone (FSH) for the treatmentprostatic carcinoma, for instance, LHRH agonists and antagonists such asgoserelin acetate and luprolide.

Signal transduction pathway inhibitors are those inhibitors, which blockor inhibit a chemical process which evokes an intracellular change. Asused herein this change is cell proliferation or differentiation. Signaltransduction inhibitors useful in the present invention includeinhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases,SH2/SH3 domain blockers, serine/threonine kinases, phosphotidylinositol-3 kinases, myo-inositol signaling, and Ras oncogenes.

Several protein tyrosine kinases catalyse the phosphorylation ofspecific tyrosyl residues in various proteins involved in the regulationof cell growth. Such protein tyrosine kinases can be broadly classifiedas receptor or non-receptor kinases.

Receptor tyrosine kinases are transmembrane proteins having anextracellular ligand binding domain, a transmembrane domain, and atyrosine kinase domain. Receptor tyrosine kinases are involved in theregulation of cell growth and are generally termed growth factorreceptors. Inappropriate or uncontrolled activation of many of thesekinases, i.e. aberrant kinase growth factor receptor activity, forexample by over-expression or mutation, has been shown to result inuncontrolled cell growth. Accordingly, the aberrant activity of suchkinases has been linked to malignant tissue growth. Consequently,inhibitors of such kinases could provide cancer treatment methods.Growth factor receptors include, for example, epidermal growth factorreceptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2,erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosinekinase with immunoglobulin-like and epidermal growth factor homologydomains (TIE-2), insulin growth factor-I (IGFI) receptor, macrophagecolony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growthfactor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin(eph) receptors, and the RET protooncogene. Several inhibitors of growthreceptors are under development and include ligand antagonists,antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides.Growth factor receptors and agents that inhibit growth factor receptorfunction are described, for instance, in Kath, John C., Exp. Opin. Ther.Patents (2000) 10(6):803-818; Shawver et al DDT Vol 2, No. 2 Feb. 1997;and Lofts, F. J. et al, “Growth factor receptors as targets”, NewMolecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr,David, CRC press 1994, London.

Tyrosine kinases, which are not growth factor receptor kinases aretermed non-receptor tyrosine kinases. Non-receptor tyrosine kinasesuseful in the present invention, which are targets or potential targetsof anti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focaladhesion kinase), Brutons tyrosine kinase, and Bcr-Abl. Suchnon-receptor kinases and agents which inhibit non-receptor tyrosinekinase function are described in Sinh, S, and Corey, S. J., (1999)Journal of Hematotherapy and Stem Cell Research 8 (5): 465-80; andBolen, J. B., Brugge, J. S., (1997) Annual review of Immunology. 15:371-404.

SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domainbinding in a variety of enzymes or adaptor proteins including, PI3-K p85subunit, Src family kinases, adaptor molecules (Shc, Crk, Nck, Grb2) andRas-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussedin Smithgall, T. E. (1995), Journal of Pharmacological and ToxicologicalMethods. 34(3) 125-32.

Inhibitors of Serine/Threonine Kinases including MAP kinase cascadeblockers which include blockers of Raf kinases (rafk), Mitogen orExtracellular Regulated Kinase (MEKs), and Extracellular RegulatedKinases (ERKs); and Protein kinase C family member blockers includingblockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta).IkB kinase family (IKKa, IKKb), PKB family kinases, AKT kinase familymembers, and TGF beta receptor kinases. Such Serine/Threonine kinasesand inhibitors thereof are described in Yamamoto, T., Taya, S.,Kaibuchi, K., (1999), Journal of Biochemistry. 126 (5) 799-803; Brodt,P, Samani, A., and Navab, R. (2000), Biochemical Pharmacology, 60.1101-1107; Massague, J., Weis-Garcia, F. (1996) Cancer Surveys.27:41-64; Philip, P. A., and Harris, A. L. (1995), Cancer Treatment andResearch. 78: 3-27, Lackey, K. et al Bioorganic and Medicinal ChemistryLetters, (10), 2000, 223-226; U.S. Pat. No. 6,268,391; andMartinez-Iacaci, L., et al, Int. J. Cancer (2000), 88(1), 44-52.

Inhibitors of Phosphotidyl inositol-3 Kinase family members includingblockers of PI3-kinase, ATM, DNA-PK, and Ku are also useful in thepresent invention. Such kinases are discussed in Abraham, R. T. (1996),Current Opinion in Immunology. 8 (3) 412-8; Canman, C. E., Lim, D. S.(1998), Oncogene 17 (25) 3301-3308; Jackson, S. P. (1997), InternationalJournal of Biochemistry and Cell Biology. 29 (7):935-8; and Zhong, H. etal, Cancer res, (2000) 60(6), 1541-1545.

Also useful in the present invention are Myo-inositol signalinginhibitors such as phospholipase C blockers and Myoinositol analogues.Such signal inhibitors are described in Powis, G., and Kozikowski A.,(1994) New Molecular Targets for Cancer Chemotherapy ed., Paul Workmanand David Kerr, CRC press 1994, London.

Another group of signal transduction pathway inhibitors are inhibitorsof Ras Oncogene. Such inhibitors include inhibitors offarnesyltransferase, geranyl-geranyl transferase, and CAAX proteases aswell as anti-sense oligonucleotides, ribozymes and immunotherapy. Suchinhibitors have been shown to block ras activation in cells containingwild type mutant ras, thereby acting as antiproliferation agents. Rasoncogene inhibition is discussed in Scharovsky, O. G., Rozados, V. R.,Gervasoni, S. I. Matar, P. (2000), Journal of Biomedical Science. 7(4)292-8; Ashby, M. N. (1998), Current Opinion in Lipidology. 9 (2) 99-102;and BioChem. Biophys. Acta, (19899) 1423(3):19-30.

As mentioned above, antibody antagonists to receptor kinase ligandbinding may also serve as signal transduction inhibitors. This group ofsignal transduction pathway inhibitors includes the use of humanizedantibodies to the extracellular ligand binding domain of receptortyrosine kinases. For example Imclone C225 EGFR specific antibody (seeGreen, M. C. et al, Monoclonal Antibody Therapy for Solid Tumors, CancerTreat. Rev., (2000), 26(4), 269-286); Herceptin® erbB2 antibody (seeTyrosine Kinase Signalling in Breast cancer:erbB Family ReceptorTyrosine Kinases, Breast cancer Res., 2000, 2(3), 176-183); and 2CBVEGFR2 specific antibody (see Brekken, R. A. et al, Selective Inhibitionof VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumorgrowth in mice, Cancer Res. (2000) 60, 5117-5124).

Non-receptor kinase angiogenesis inhibitors may also find use in thepresent invention Inhibitors of angiogenesis related VEGFR and TIE2 arediscussed above in regard to signal transduction inhibitors (bothreceptors are receptor tyrosine kinases). Accordingly, non-receptortyrosine kinase inhibitors may be used in combination with theinhibitors of the present invention. For example, anti-VEGF antibodies,which do not recognize VEGFR (the receptor tyrosine kinase), but bind tothe ligand; small molecule inhibitors of integrin (alpha beta₃) thatwill inhibit angiogenesis; endostatin and angiostatin (non-RTK) may alsoprove useful in combination with the disclosed family inhibitors. (SeeBruns C J et al (2000), Cancer Res., 60: 2926-2935; Schreiber A B,Winkler M E, and Derynck R. (1986), Science, 232: 1250-1253; Yen L etal. (2000), Oncogene 19: 3460-3469).

Agents used in immunotherapeutic regimens may also be useful incombination with the compounds of formula (I). There are a number ofimmunologic strategies to generate an immune response against erbB2 orEGFR. These strategies are generally in the realm of tumor vaccinations.The efficacy of immunologic approaches may be greatly enhanced throughcombined inhibition of erbB2/EGFR signaling pathways using a smallmolecule inhibitor. Discussion of the immunologic/tumor vaccine approachagainst erbB2/EGFR are found in Reilly R T et al. (2000), Cancer Res.60: 3569-3576; and Chen Y, Hu D, Eling D J, Robbins J, and Kipps T J.(1998), Cancer Res. 58: 1965-1971.

Agents used in proapoptotic regimens (e.g., bcl-2 antisenseoligonucleotides) may also be used in the combination of the presentinvention. Members of the Bcl-2 family of proteins block apoptosis.Upregulation of bcl-2 has therefore been linked to chemoresistance.Studies have shown that the epidermal growth factor (EGF) stimulatesanti-apoptotic members of the bcl-2 family (i.e., mcl-1). Therefore,strategies designed to downregulate the expression of bcl-2 in tumorshave demonstrated clinical benefit and are now in Phase II/III trials,namely Genta's G3139 bcl-2 antisense oligonucleotide. Such proapoptoticstrategies using the antisense oligonucleotide strategy for bcl-2 arediscussed in Water J S et al. (2000), J. Clin. Oncol. 18: 1812-1823; andKitada S et al. (1994), Antisense Res. Dev. 4: 71-79.

Cell cycle signalling inhibitors inhibit molecules involved in thecontrol of the cell cycle. A family of protein kinases called cyclindependent kinases (CDKs) and their interaction with a family of proteinstermed cyclins controls progression through the eukaryotic cell cycle.The coordinate activation and inactivation of different cyclin/CDKcomplexes is necessary for normal progression through the cell cycle.Several inhibitors of cell cycle signalling are under development. Forinstance, examples of cyclin dependent kinases, including CDK2, CDK4,and CDK6 and inhibitors for the same are described in, for instance,Rosania et al, Exp. Opin. Ther. Patents (2000) 10(2):215-230.

In one embodiment, the cancer treatment method of the claimed inventionincludes the co-administration a compound of formula I and/or apharmaceutically acceptable salt, hydrate, solvate or pro-drug thereofand at least one anti-neoplastic agent, such as one selected from thegroup consisting of anti-microtubule agents, platinum coordinationcomplexes, alkylating agents, antibiotic agents, topoisomerase IIinhibitors, antimetabolites, topoisomerase I inhibitors, hormones andhormonal analogues, signal transduction pathway inhibitors, non-receptortyrosine kinase angiogenesis inhibitors, immunotherapeutic agents,proapoptotic agents, and cell cycle signaling inhibitors.

The pharmaceutically active compounds of the present invention areincorporated into convenient dosage forms such as capsules, tablets, orinjectable preparations. Solid or liquid pharmaceutical carriers areemployed. Solid carriers include, starch, lactose, calcium sulfatedihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia,magnesium stearate, and stearic acid. Liquid carriers include syrup,peanut oil, olive oil, saline, and water. Similarly, the carrier ordiluent may include any prolonged release material, such as glycerylmonostearate or glyceryl distearate, alone or with a wax. The amount ofsolid carrier varies widely but, preferably, will be from about 25 mg toabout 1 g per dosage unit. When a liquid carrier is used, thepreparation will be in the form of a syrup, elixir, emulsion, softgelatin capsule, sterile injectable liquid such as an ampoule, or anaqueous or nonaqueous liquid suspension.

The pharmaceutical preparations are made following conventionaltechniques of a pharmaceutical chemist involving mixing, granulating,and compressing, when necessary, for tablet forms, or mixing, fillingand dissolving the ingredients, as appropriate, to give the desired oralor parenteral products.

Doses of the presently invented pharmaceutically active compounds in apharmaceutical dosage unit as described above will be an efficacious,nontoxic quantity preferably selected from the range of 0.001-100 mg/kgof active compound, preferably 0.001-50 mg/kg. When treating a humanpatient in need of a PI3K inhibitor, the selected dose is administeredpreferably from 1-6 times daily, orally or parenterally. Preferred formsof parenteral administration include topically, rectally, transdermally,by injection and continuously by infusion. Oral dosage units for humanadministration preferably contain from 0.05 to 3500 mg of activecompound. According to one embodiment, the oral dosage for humanadministration contains 100 to 1000 mg per day. Oral administration,which uses lower dosages is preferred. Parenteral administration, athigh dosages, however, also can be used when safe and convenient for thepatient.

Optimal dosages to be administered may be readily determined by thoseskilled in the art, and will vary with the particular PI3 kinaseinhibitor in use, the strength of the preparation, the mode ofadministration, and the advancement of the disease condition. Additionalfactors depending on the particular patient being treated will result ina need to adjust dosages, including patient age, weight, diet, and timeof administration. Exemplary dosages include oral formulationsequivalent to 10 mg, 25 mg, and 100 mg of the compound of formula (I),to be administered alone, in multiples, or in combination. Anotherexemplary dosage includes oral formulations of thetris(hydroxymethyl)aminomethane salt of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid equivalent to 10 mg, 25 mg, or 100 mg of the free base of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid.

The method of this invention of inducing PI3 kinase inhibitory activityin mammals, including humans, comprises administering to a subject inneed of such activity an effective PI3 kinase modulating/inhibitingamount of a pharmaceutically active compound of the present invention.

The invention also provides for the use of a compound of Formula (I) inthe manufacture of a medicament for use as a PI3 kinase inhibitor.

The invention also provides for the use of a compound of Formula (I) inthe manufacture of a medicament for use in therapy.

The invention also provides for the use of a compound of Formula (I) inthe manufacture of a medicament for use in treating autoimmunedisorders, inflammatory diseases, cardiovascular diseases,neurodegenerative diseases, allergy, asthma, pancreatitis, multiorganfailure, kidney diseases, platelet aggregation, cancer, sperm motility,transplantation rejection, graft rejection and lung injuries.

The invention also provides for a pharmaceutical composition for use asa PI3 inhibitor which comprises a compound of Formula (I) and apharmaceutically acceptable carrier.

The invention also provides for a pharmaceutical composition for use inthe treatment of autoimmune disorders, inflammatory diseases,cardiovascular diseases, neurodegenerative diseases, allergy, asthma,pancreatitis, multiorgan failure, kidney diseases, platelet aggregation,cancer, sperm motility, transplantation rejection, graft rejection andlung injuries, which comprises a compound of Formula (I) and apharmaceutically acceptable carrier.

In addition, the pharmaceutically active compounds of the presentinvention can be co-administered with further active ingredients,including compounds known to have utility when used in combination witha PI3 kinase inhibitor.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present invention toits fullest extent. The following examples are, therefore, to beconstrued as merely illustrative and not a limitation of the scope ofthe present invention in any way.

EXPERIMENTAL PROCEDURES

Compounds of Formula (I) may be prepared using the general schemesI-VII, as described below.

Scheme I (R2=Me)

2,6-dinitro aniline 1 can be brominated with bromine in acetic acid toprovide 4-bromo-2,6-dinitroaniline 2 that can be reduced to the di-aminonitro benzene 3 with (NH₄ ⁺)₂S. Subsequent reaction of 3 with2,4-pentanedione in the presence of strong acid at reflux temperatures,in an alcoholic solvent, affords nitrobenzimidazole 4. Alkylation toafford substituted benzimidazole 5 can be accomplished with a suitablysubstituted alkyl halide with a base, such as K₂CO₃, in a polar aproticsolvent, such as DMF. Palladium-catalyzed displacement of the aromaticbromine with morpholine can then afford substituted nitrobenzimidazole 6which can then be reduced to the amino benzimidazole 7. Aminobenzimidazole 7 can then be converted into hydroxyl analog 8,sulfonamide 9, amide 10, and halo analog 11, using standard organicmanipulations.

2,6-dinitro aniline 1 can be brominated with bromine in acetic acid toprovide 4-bromo-2,6-dinitroaniline 2 that can be reduced to the di-aminonitro benzene 3 with (NH₄ ⁺)₂S. Subsequent reaction of 3 with acarboxylic acid in the presence of strong acid at elevated temperaturesaffords nitrobenzimidazole 4. Alkylation to afford substitutedbenzimidazole 5 can be accomplished with a suitably substituted alkylhalide with a base, such as K₂CO₃, in a polar aprotic solvent, such asDMF. Palladium-catalyzed displacement of the aromatic bromine withmorpholine can then afford substituted nitrobenzimidazole 6 which canthen be reduced to the amino benzimidazole 7 Amino benzimidazole 7 canthen be converted into hydroxyl analog 8, sulfonamide 9, amide 10, andhalo analog 11, using standard organic manipulations.

2-amino-3-nitrophenol 1 can be methylated with MeI and K₂CO₃ in DMF toafford methoxy nitro aniline 2. Bromination, with bromine in aceticacid, followed by acetylation with acetic anhydride in acetic acid andsulfuric acid, can provide intermediate 4. Palladium-catalyzeddisplacement of the aromatic bromide with morpholine can then affordintermediate 5. Iron-induced nitro reduction followed by ring closurecan then provide benzimidazole 6 that can be alkylated with a suitablysubstituted alkyl bromide using a base, such as K₂CO₃, in a polaraprotic solvent such as DMF, to afford final products 7.

Aminobenzimidazole 1 can be converted to bromobenzimidazole 2 usingsodium nitrite with NaBr in aqueous HBr. Palladium catalyzed couplingwith an aryl boronic acid in the presence of a suitable phosphine withan inorganic base in a polar non-protic solvent can then provide finalsubstituted benzimidazoles 3. Het includes 2-, 3-furanyls, and1,3-thiozols.

Palladium catalyzed carbonylaton of bromo-benzimidazole 1 can beaccomplished by bubbling carbon-monoxide gas in methanol withtriethylamine to provide methyl ester 2. Ester hydrolysis can then beaccomplished with lithium hydroxide in THF/water to provide finalproduct benzimidazole acid 3.

Palladium catalyzed cyanation of bromo-benzimidazole 1 can beaccomplished with zinc cyanide in DMF to provide benzimidazole nitrile2. The nitrile can be converted to the primary carboxamide with KOH andperoxide in THF to provide amide 3. Treatment of the carboxamide 3 withDMF-DMA can provide intermediate 4 that can then be cyclized to triazoleanalogs 5 with hydrazine in acetic acid.

Amination of 5-chloro-2-nitrobenzoic acid with O-methyl hydroxylamineand t-butoxide in the presence of copper acetate can provide3-amino-5-chloro-2-nitrobenzoic acid 2. Esterification can beaccomplished with methanol and sulfuric acid to provide methyl ester 3that can be reacted with morpholine in DMF with K₂CO₃ to provide phenylmorpholine analog 4. Nitro reduction can be accomplished using a varietyof metal reductions to provide diamine 5. Condensation of 5 with avariety of carboxylic acids can provide benzimidazole methylester 6 thatcan be further converted to final products 7 (R1=CO₂Me, CO₂H, CONH₂, CN,triazole, tetrazole) after alkylation with an alkyl halide, followed bystandard organic manipulations as previously described.

Preparation of 5-bromo-2-methyl-7-nitro-1H-benzimidazole a)4-bromo-2,6-dinitrobenzenamine

A stirred suspension of 2,6-dinitroaniline (5 g, 27.3 mmol) in glacialacetic acid (50 mL) was added bromine (1.5 mL, 30 mmol) dropwise andheated at 120° C. for 2 h. After cooling to ambient temperature, theresultant mixture was poured into water (50 mL). The precipitate solidwas collected by filtration and washed with water then dried in-vacuo.The solid was re-dissolved in EtOAC, washed with water and saturatedbrine. The organic layer was collected and concentrated in-vacuo to givethe desired product (6.88 g, 95%). ¹H NMR (300 MHz, DMSO-d₆) δ ppm 8.37(br s, 2H), 8.58 (s, 2H).

b) 5-bromo-3-nitrobenzene-1,2-diamine

4-bromo-2,6-dinitrobenzenamine was dissolved in EtOH (50 mL) and (NH₄)₂S(2.2 mL) was added to the mixture. The reaction mixture was heated to90° C. for 1 h. TLC showed a new compound and some remaining startingmaterial remained. Additionally, another batch (NH₄)₂S (2.5 mL) wasadded. After 1 h, TLC analysis showed little starting material remained.The reaction mixture was concentrated to give a deep red solid. It wasthen purified by silica gel chromatography eluted with DCM to afford thedesired product as a red solid (578 mg, 50%). ¹H NMR (300 MHz, CDCl₃) δppm 3.50 (br s, 2H), 5.93 (br s, 2H), 7.04 (d, 1H, J=1.8 Hz), 7.87 (d,1H, J=1.8 Hz); LC-MS: m/e=232 [M+1]⁺.

c) 6-bromo-2-methyl-4-nitro-1H-benzo[d]imidazole

A mixture of 5-bromo-3-nitrobenzene-1,2-diamine (464 mg) andpentane-2,4-dione (400 mg) in EtOH (27 mL) and 5 N HCl (7.4 mL) wasrefluxed for 3 h. The mixture was cooled to room temperature and thesolvent was removed in-vacuo. The residue was dissolved in EtOAc andwashed with aqueous NaHCO₃ solution and brine. The organic layer wasconcentrated to afford the desired product as a solid (460 mg, 90%). ¹HNMR (300 MHz, CDCl₃) δ ppm 2.73 (s, 3H), 8.11 (d, 1H, J=1.8 Hz), 8.24(d, 1H, J=1.8 Hz), 10.20 (s, 1H, s); LC-MS: m/e=256 [M+1]⁺

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-amineandN-(2-Methyl-6-morpholin-4-yl-1-naphthalen-1-ylmethyl-1H-benzoimidazol-4-yl)-acetamidea)6-bromo-2-methyl-1-(naphthalen-1-ylmethyl)-4-nitro-1H-benzo[d]imidazole

A mixture of 6-bromo-2-methyl-4-nitro-1H-benzo[d]imidazole (preparedfollowing the same procedure as Example 1) (3 g),1-(bromomethyl)naphthalene (2.85 g) and K₂CO₃ (3.23 g) in DMF (100 mL)was stirred at 80° C. overnight. It was cooled to room temperature andfiltered. The filtrate was then poured into water. It was then filteredto afford a solid and the solid was washed with water and then driedin-vacuo to afford the desired product (4.63 g, 100%). ¹H NMR (300 MHz,DMSO-d₆) δ ppm 2.54 (s, 3H), 6.16 (s, 2H), 6.32 (d, 1H, J=7.5 Hz), 7.33(t, 1H, J=7.5 Hz), 7.61-7.72 (m, 2H), 7.87 (d, 1H, J=7.5 Hz), 8.01 (d,1H, J=7.5 Hz), 8.14 (d, 1H, J=1.8 Hz), 8.19 (d, 1H, J=7.5 Hz), 8.28 (d,1H, J=1.8 Hz); LC-MS: m/e=296 [M+1]⁺.

b)4-(2-methyl-3-(naphthalen-1-ylmethyl)-7-nitro-3H-benzo[d]imidazol-5-yl)morpholine

A mixture of6-bromo-2-methyl-1-(naphthalen-1-ylmethyl)-4-nitro-1H-benzo[d]imidazole(4.63 g), morpholine (3.05 g), Pd₂(dba)₃ (1.05 g), Cs₂CO₃ (5.72 g) andX-Phos (1.09 g) in dioxane (100 mL) was degassed with nitrogen and thenstirred at 80° C. overnight. The mixture was cooled to room temperatureand the solvent was removed in-vacuo. The residue was then purified bysilica gel chromatography eluted with EtOAc:Petrolium ether=1:1 toafford the desired product as a yellow solid (2.8 g, 60%). ¹H NMR (300MHz, DMSO-d6) δ ppm 2.46 (s, 3H), 3.12 (t, 4H, J=4.8 Hz). 3.70 (t, 4H,J=4.8 Hz), 6.09 (s, 2H), 6.31 (d, 1H, J=7.5 Hz), 7.34 (t, 1H, J=7.5 Hz),7.53 (d, 1H, J=2.1 Hz), 7.62-7.70 (m, 3H), 7.86 (d, 1H, J=8.1 Hz), 8.01(d, 1H, J=7.5 Hz), 8.23 (d, 1H, J=8.1 Hz); LC-MS: m/e=403 [M+1]+.

c)2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-amineandN-(2-Methyl-6-morpholin-4-yl-1-naphthalen-1-ylmethyl-1H-benzoimidazol-4-yl)-acetamide

To a refluxing mixture of4-(2-methyl-3-(naphthalen-1-ylmethyl)-7-nitro-3H-benzo[d]imidazol-5-yl)morpholine(804 mg) in HOAc (50 mL) was added iron powder (336 mg) and the mixturewas continued to reflux for 3 h. The mixture was cooled to roomtemperature and HOAc was removed in-vacuo. The residue was thenneutralized with aqueous NaHCO₃ solution. It was extracted with DCM andthe organic layer was washed with brine, dried over anhydrous Na₂SO₄,filtered and concentrated in-vacuo. The residue was purified by silicagel chromatography eluted with MeOH:DCM=1:30 to afford Example 2 (350mg, 47%) and Example 3 (350 mg, 42%). Example 2 ¹H NMR (300 MHz,DMSO-d₆) δ ppm 2.33 (s, 3H), 2.91 (t, 4H J=4.8 Hz), 3.64 (t, 4H J=4.8Hz), 5.15 (br s, 2H), 5.83 (s, 2H), 6.10 (d, 1H, J=2.1 Hz), 6.12 (d, 1H,J=2.1 Hz), 6.38 (d, 1H, J=7.5 Hz), 7.34 (t, 1H, J=7.5 Hz), 7.58-7.68 (m,2H), 7.84 (d, 1H, J=8.4 Hz), 8.01 (d, 1H, J=7.5 Hz), 8.23 (d, 1H, J=8.4Hz); LC-MS: m/e=373 [M+1]+; Example 3 ¹H NMR (300 MHz, DMSO-d₆) δ ppm2.18 (s, 3H), 2.40 (s, 3H, s), 2.96 (t, 4H, J=4.8 Hz), 3.67 (t, 4H,J=4.8 Hz), 5.95 (s, 2H), 6.34 (d, 1H, J=7.5 Hz), 6.68 (s, 1H), 7.34 (t,1H, J=7.5 Hz), 7.59-7.70 (m, 2H), 7.76 (s, 1H), 7.85 (d, 1H, J=7.5 Hz),8.01 (d, 1H, J=7.5 Hz), 8.23 (d, 1H, J=7.5 Hz), 9.81 (s, 1H); LC-MS:m/e=415 [M+1]+

Preparation ofN-[2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-yl]methanesulfonamidea)2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-amine

A mixture of4-(2-methyl-3-(naphthalen-1-ylmethyl)-7-nitro-3H-benzo[d]imidazol-5-yl)morpholine(804 mg), prepared as described in Example 2, iron powder (168 mg) andFeSO₄ (84 mg) in ethanol (30 mL) and H₂O (30 mL) was stirred at refluxtemperature overnight. The mixture was cooled to room temperature andthe solvent was removed in-vacuo. The residue was dissolved in DCM andfiltered. The filtrate was then washed with brine, dried over anhydrousNa₂SO₄, filtered and concentrated in-vacuo to afford the desired productas a solid (720 mg, 97%). ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.33 (s, 3H),2.91 (t, 4H J=4.8 Hz), 3.64 (t, 4H J=4.8 Hz), 5.15 (br s, 2H), 5.83 (s,2H), 6.10 (d, 1H, J=2.1 Hz), 6.12 (d, 1H, J=2.1 Hz), 6.38 (d, 1H, J=7.5Hz), 7.34 (t, 1H, J=7.5 Hz), 7.58-7.68 (m, 2H), 7.84 (d, 1H, J=8.4 Hz),8.01 (d, 1H, J=7.5 Hz), 8.23 (d, 1H, J=8.4 Hz); LC-MS: m/e=373 [M+1]+

b)N-(2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-yl)methanesulfonamide

To a solution of2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-amine(186 mg), Et₃N (0.15 mL) and DCM (20 mL) was added a solution ofmethanesulfonyl chloride (69 mg) in DCM at 0° C. and then the mixturewas stirred at room temperature for 1 h. The mixture was diluted withDCM and washed with brine, dried over anhydrous Na₂SO₄, filtered andconcentrated in-vacuo. The residue was then purified by silica gelchromatography eluted with MeOH:DCM=1:30 to afford the desired productas a solid (180 mg, 80%). ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.40 (s, 3H),2.98 (t, 4H, J=4.8 Hz), 3.21 (s, 3H), 3.68 (t, 4H, J=4.8 Hz), 5.96 (s,2H), 6.37 (d, 1H, J=8.1 Hz), 6.80 (s, 2H), 7.35 (t, 1H, J=8.1 Hz),7.60-7.71 (m, 2H), 7.85 (d, 1H, J=8.1 Hz), 8.01 (d, 1H, J=8.1 Hz), 8.24(d, 1H, J=8.1 Hz), 9.49 (br s, 1H); LC-MS: m/e=451 [M+1]+.

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-ola)2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-amine

TiCl₃ (19.7 mL) was added to a solution of4-(2-methyl-3-(naphthalen-1-ylmethyl)-7-nitro-3H-benzo[d]imidazol-5-yl)morpholine(1.82 g), prepared following the same procedure as in Example 4, andNH₄OAc (4.85 g) in MeOH (150 mL). After stirring for 7 min at roomtemperature, TLC showed no starting material remaining. The pH of themixture was made basic by adding aqueous Na₂CO₃ solution. The solventwas removed under reduced pressure and the residue was extracted withDCM (250 mL×2). The combined organic layers were washed with brine,dried over anhydrous Na₂SO₄, filtered and concentrated in-vacuo toafford the desired product as a white solid (1.52 g, 91%). ¹H NMR (300MHz, CDCl₃) δ ppm 2.47 (s, 3H), 3.02 (t, 4H, J=4.8 Hz), 3.78 (t, 4H,J=4.8 Hz), 4.30 (s, 2H), 5.68 (s, 2H), 6.05 (d, 1H, J=1.8 Hz), 6.25 (d,1H, J=1.8 Hz), 6.56 (d, 1H, J=7.5 Hz), 7.27 (t, 1H, J=7.5 Hz), 7.55-7.66(m, 2H), 7.77 (d, 1H, J=8.1 Hz), 7.93 (d, 1H, J=8.1 Hz), 8.05 (d, 1H,J=8.1 Hz); LC-MS: m/e=373 [M+1]+.

b)2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-ol

To a solution of2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-amine(842 mg) in H₂O (20 mL), MeOH (1 mL) and conc. H₂SO₄ (3 mL) was addedaqueous NaNO₂ (344 mg) solution drop-wise at 0° C. The mixture wasstirred at 0° C. for 15 min and then stirred at reflux temperature for 1h. The mixture was cooled to room temperature and the pH neutralizedwith aqueous NaHCO₃ solution. It was extracted with DCM (100 mL×3). Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄, filtered and concentrated in-vacuo. The residue was thenpurified by silica gel chromatography eluted with MeOH:DCM=1:60 and thenby Prep-HPLC to afford crude desired product LC-MS: m/e=374 [M+1]+containing an impurity that was removed by the two-step sequencedescribed below.

c)4-{[(1,1-dimethylethyl)(diphenyl)silyl]oxy}-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole

A mixture of crude2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-ol(200 mg), imidazole (73 mg) and TBDPSCl (162 mg) in dry DCM (30 mL) wasstirred at room temperature for 1 h. LCMS analysis showed desiredproduct so the solvent was removed in-vacuo. The residue was purified bysilica gel chromatography eluted with EtOAc:petroleum ether=1:2 toafford the TBDP ether desired product as a white solid (260 mg, 79%). ¹HNMR (300 MHz, CDCl₃) δ ppm 1.24 (s, 9H), 2.53-2.56 (m, 7H), 3.55 (t, 4H,J=4.8 Hz), 5.69 (s, 2H), 5.92 (d, 1H, J=1.8 Hz), 6.12 (d, 1H, J=1.8 Hz),6.55 (d, 1H, J=7.5 Hz), 7.25-7.45 (m, 7H), 7.56-7.67 (m, 2H), 7.78 (d,1H, J=8.7 Hz), 7.84-7.87 (m 4H), 7.94 (d, 1H, J=7.5 Hz), 8.06 (d, 1H,J=8.7 Hz); LC-MS: m/e=612 [M+1]+.

d)2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-ol

To a solution of4-{[(1,1-dimethylethyl)(diphenyl)silyl]oxy}-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazolein THF (50 mL) was added TBAF (0.64 mL, 1 mol/L) at room temperature andthe mixture was stirred for 1 h. TLC showed consumption of startingmaterial. The solvent was removed in-vacuo and the residue was purifiedby silica gel chromatography eluted with MeOH:DCM=1:60 to afford thedesired product as a white solid (150 mg, 94%). ¹H NMR (300 MHz,DMSO-d₆) δ ppm 2.36 (s, 3H), 2.94 (t, 4H, J=4.8 Hz), 3.65 (t, 4H, J=4.8Hz), 5.87 (s, 2H), 6.30 (d, 1H, J=1.8 Hz), 6.35 (d, 1H, J=1.8 Hz), 6.39(d, 1H, J=7.5 Hz), 7.35 (t, 1H, J=7.5 Hz), 7.60-7.71 (m, 2H), 7.86 (d,1H, J=8.4 Hz), 8.01 (d, 1H, J=7.5 Hz), 8.23 (d, 1H, J=8.4 Hz); LC-MS:m/e=374[M+1]+

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-methyl-4-(methyloxy)-6-(4-morpholinyl)-1H-benzimidazolea) 2-methoxy-6-nitrobenzenamine

To a mixture of 2-amino-3-nitrophenol (19.25 g) and K₂CO₃ (19 g) in DMF(100 mL) was added MeI (11 mL) at room temperature and the mixture wasstirred over night and then poured into water. The resulting precipitatewas collected by filtration and the solid was washed with water toafford the desired product (19 g, 90%). ¹H NMR (300 MHz, CDCl₃) δ ppm3.92 (s, 1H), 6.43 (br s, 1H), 6.61 (dd, 1H, J=7.5, 9.0 Hz), 6.89 (dd,1H, J=0.9, 7.5 Hz), 7.73 (dd, 1H, J=0.9, 9.0 Hz); LC-MS: m/e=169 [M+1]+

b) 4-bromo-2-methoxy-6-nitrobenzenamine

NaOAc (17.6 g) and Br₂ (6.76 mL) was added to a solution of2-methoxy-6-nitrobenzenamine (21.74 g) in HOAc (250 mL). The mixture wasstirred at room temperature for 20 min. The resulting precipitate wasfiltered, washed with water and dried in-vacuo to afford the desiredproduct as a yellow solid (26.43 g, 83%). ¹H NMR (300 MHz, DMSO-d₆) δ3.91 (s, 3H), 7.18 (d, 1H, J=1.8 Hz), 7.70 (d, 1H, J=1.8 Hz); LC-MS:m/e=247 [M+1]+

c) N-(4-bromo-2-methoxy-6-nitrophenyl)acetamide

To a solution of 4-bromo-2-methoxy-6-nitrobenzenamine (27.85 g) in HOAc(150 mL) and Ac₂O (17 mL) was added conc. H₂SO₄ at 70° C. and themixture was stirred at 70° C. for 30 min and kept at rt overnight. Theformed precipitate was collected by filtration and washed with hexane toafford the desired product as a light yellow solid (24.45 g, 75%). ¹HNMR (300 MHz, DMSO-d6) δ ppm 2.01 (s, 3H), 3.92 (s, 3H), 7.61 (d, 1H,J=1.8 Hz), 7.65 (d, 1H, J=1.8 Hz), 9.91 (s, 1H); LC-MS: m/e=289 [M+1]+

d) N-(2-methoxy-4-morpholino-6-nitrophenyl)acetamide

A mixture of N-(4-bromo-2-methoxy-6-nitrophenyl)acetamide (2.89 g),morpholine (2.61 g), BINAP (1.21 g) and t-BuOK (1.53 g) in dioxane (50mL) was degassed with N₂ and the mixture was stirred at 110° C. in asealed tube overnight. It was cooled to room temperature and filtered.The filtrate was concentrated in-vacuo. The residue was purified bysilica gel chromatography eluted with MeOH/DCM=1/50 to afford thedesired product (1.03 g, 35%) ¹H NMR (300 MHz, CDCl₃) δ ppm 2.15 (s,1H), 3.18 (t, 4H, J=4.8 Hz), 3.85 (t, 4H, J=4.8 Hz), 3.88 (s, 3H), 6.63(d, 1H, J=2.7 Hz), 6.96 (d, 1H, J=2.7 Hz).

e) 4-(7-methoxy-2-methyl-3H-benzo[d]imidazol-5-yl)morpholine

To a refluxing solution of combined batches ofN-(2-methoxy-4-morpholino-6-nitrophenyl)acetamide (2.06 g) in HOAc (60mL) was added iron powder (1.18 g) and the mixture was stirred at refluxtemperature overnight. It was cooled to room temperature and filtered.The filtrate was concentrated in-vacuo and the residue was washed withEtOAc:petroleum ether=1:1 to afford crude product as a solid (1.73 g,100%). ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.56 (s, 3H), 3.12 (t, 4H, J=4.8Hz), 3.88 (t, 4H, J=4.8 Hz), 3.94 (s, 3H), 6.39 (s, 1H), 6.62 (s, 1H);LC-MS: m/e=248 [M+1]+

f)1-[(2,3-dichlorophenyl)methyl]-2-methyl-4-(methyloxy)-6-(4-morpholinyl)-1H-benzimidazole

A mixture of 18 (1.73 g), 1-(bromomethyl)-2,3-dichlorobenzene (1.68 g)and K₂CO₃ (1.93 g) in DMF (50 mL) was stirred at 80° C. for 72 h. Themixture was cooled to room temperature and poured into water. It wasextracted with EtOAc and the organic layer was washed with brine, driedover anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residuewas then purified by silica gel chromatography eluted with 100% EtOAcand then MeOH:DCM=1:30 to afford the desired product as a white solid(360 mg, 9%) ¹H NMR (CDCl₃, TMS, 300 MHz) δ ppm 2.48 (s, 3H), 3.11 (t,4H, J=4.8 Hz), 3.85 (t, 4H, J=4.8 Hz), 4.02 (s, 3H), 5.31 (s, 2H), 6.19(d, 1H, J=1.8 Hz), 6.30 (d, 1H, J=7.5 Hz), 6.42 (d, 1H, J=1.8 Hz), 7.03(t, 1H, J=7.5 Hz), 7.41 (d, 1H, J=7.5 Hz); LC-MS: m/e=406 [M+1]+

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazol-4-aminea) 6-bromo-1-(2,3-dichlorobenzyl)-2-methyl-4-nitro-1H-benzo[d]imidazole

A mixture of Example 1 (1.17 g) (prepared as described previouslydescribed), 1-(bromomethyl)-2,3-dichlorobenzene (1.19 g) and K₂CO₃ (1.27g) in DMF (80 mL) was stirred at 80° C. for 3 h. When TLC showed nostarting material, the mixture was cooled to room temperature andfiltered. The filtrate was then poured into water. It was then filteredto afford a solid and the solid was washed with water and then driedin-vacuo to afford the desired product (1.59 g, 83%) ¹H NMR (300 MHz,CDCl₃) δ ppm 2.67 (s, 3H), 5.45 (m, 2H), 6.24 (t, 1H, J=7.8 Hz), 7.10(t, 1H, J=7.8 Hz), 7.47 (t, 1H, J=7.8 Hz), 7.59 (d, 1H, J=1.8 Hz), 8.24(d, 1H, J=1.8 Hz); LC-MS: m/e=416 [M+1]+.

b)4-(3-(2,3-dichlorobenzyl)-2-methyl-7-nitro-3H-benzo[d]imidazol-5-yl)morpholine

A mixture of6-bromo-1-(2,3-dichlorobenzyl)-2-methyl-4-nitro-1H-benzo[d]imidazole(1.69 g), morpholine (1.07 g), Pd₂(dba)₃ (376 mg), Cs₂CO₃ (2 g) andX-Phos (383 mg) in dioxane (80 mL) was degassed with nitrogen and thenstirred at 80° C. for 3 h. When TLC showed complete consumption ofstarting material, the mixture was cooled to room temperature and thesolvent was removed in-vacuo. The remaining residue was extracted withEtOAc. The organic layer was washed with brine, dried over anhydrousNa₂SO₄, filtered and concentrated in-vacuo. The residue was thenpurified by silica gel chromatography eluted with EtOAc:Petroliumether=1:1 to afford the desired product as a yellow solid (831 mg, 48%).¹H NMR (300 MHz, CDCl₃) δ ppm 2.62 (s, 3H), 3.18 (t, 4H, J=4.8 Hz), 3.87(t, 4H, J=4.8 Hz), 5.41 (s, 2H), 6.28 (d, 1H, J=7.8 Hz), 6.86 (d, 1H,J=2.4 Hz), 7.08 (t, 1H, J=7.8 Hz), 7.46 (d, 1H, J=7.8 Hz), 7.79 (d, 1H,J=2.4 Hz); LC-MS: m/e=421 [M+1]+

c)1-[2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazol-4-amine

A mixture of4-(3-(2,3-dichlorobenzyl)-2-methyl-7-nitro-3H-benzo[d]imidazol-5-yl)morpholine(210 mg), iron powder (56 mg) and FeSO₄ (152 mg) in ethanol (25 mL) andH₂O (25 mL) was stirred at reflux temperature for 3 h. When TLC showedconsumption of all starting material, the mixture was cooled to roomtemperature and filtered. The filtrate was concentrated in-vacuo and theresidue was then purified by silica gel chromatography eluted withMeOH:DCM:NH₃.H₂O=1:60:0.5% to afford the desired product as a yellowsolid (137 mg, 70%). ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.32 (s, 1H), 2.94(t, 4H, J=4.8 Hz), 3.68 (t, 4H, J=4.8 Hz), 5.16 (br, s, 2H), 5.40 (s,2H), 6.09 (d, 1H, J=1.8 Hz), 6.13 (d, 1H, J=1.8 Hz), 6.32 (dd, 1H,J=1.5, 7.5 Hz), 7.25 (t, 1H, J=7.5 Hz), 7.58 (dd, 1H, J=1.5, 7.5 Hz);LC-MS: m/e=391 [M+1]+.

Preparation ofN-[1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazol-4-yl]methanesulfonamide

To a solution of Example 7 (82 mg) and Et₃N (42 mg) and dry DCM (20 mL)was added a solution of methanesulfonyl chloride (40 mg) in DCM at 0° C.and then the mixture was stirred at room temperature for 30 min. TLCshowed no starting material remaining and some di-mesylated product wasdetected by LC-MS: m/e=547 [M+1]+. The solvent was removed in vacuo, THF(10 mL) and 2N aqueous NaOH solution (10 mL) was added. The mixture wasstirred at room temperature for 2 h. The desired product was detected asthe main product on LC-MS. It was extracted with DCM (75 mL×2) and thecombined organic layers were concentrated in-vacuo. The residue waspurified by silica gel chromatography eluted withMeOH:DCM:NH₃.H₂O=1:60:0.5% to afford the desired product as a yellowsolid (25 mg, 26%). ¹H NMR (300 MHz, DMSO-d₆) δ 2.40 ppm (s, 1H),3.03-3.04 (m, 4H), 3.09 (s, 3H), 3.70-3.75 (m, 4H), 5.5-5.56 (m, 2H),6.31-6.34 (m, 1H), 6.80-6.82 (m, 2H), 7.25-7.30 (m, 1H), 7.60-7.62 (m,1H), 9.50 (s, 1H); LC-MS: m/e=469 [M+1]+

Preparation ofN-[1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazol-4-yl]acetamide

To a solution of Example 7 (78 mg) and Et₃N (30 mg) in dry DCM (30 mL)was added a solution of Ac₂O (20 mg) in DCM at 0° C. and then themixture was stirred at room temperature for 2 h. The mixture was thenstirred at reflux temperature for until TLC showed no starting material.The mixture was cooled to room temperature and diluted with DCM (150 mL)and washed with brine (100 mL×2). The organic layer was concentratedin-vacuo and the residue was purified by silica gel chromatographyeluted with MeOH:DCM=1:60 and then by Prep-HPLC to afford the desiredproduct as a yellow solid (21 mg, 24%). ¹H NMR (300 MHz, CDCl₃) δ ppm2.28 (s, 3H), 2.48 (s, 3H), 3.14 (t, 4H, J=4.8 Hz), 3.84 (t, 4H, J=4.8Hz), 5.33 (s, 2H), 6.28-6.30 (m, 2H), 7.05 (t, 1H, J=8.1 Hz), 7.42 (dd,1H, J=1.2 Hz, J=8.1 Hz), 8.08 (d, 1H, J=1.8 Hz), 8.27 (br s, 1H); LC-MS:m/e=433 [M+1]+.

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazol-4-ol

The titled compound was prepared following the same procedure as Example5 replacing 1-(bromomethyl)naphthalene with1-(bromomethyl)-2,3-dichlorobenzene. (130 mg, 69%). ¹H NMR (300 MHz,DMSO-d₆) δ ppm 2.35 (s, 3H), 2.97 (t, 4H, J=4.8 Hz), 3.68 (t, 4H, J=4.8Hz), 5.44 (s, 2H), 6.24 (d, 1H, J=2.1 Hz), 6.31 (dd, 1H, J=1.2, 7.8 Hz),6.38 (d, 1H, J=2.1 Hz), 7.26 (t, 1H, J=7.8 Hz), 7.59 (dd, 1H, J=1.2, 7.8Hz), 9.61 (s, 1H); LC-MS: m/e=392[M+1]+.

Preparation of2-(1-methylethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-ola) 6-bromo-2-isopropyl-4-nitro-1H-benzo[d]imidazole

A mixture of 5-bromo-3-nitrobenzene-1,2-diamine (prepared following thesame procedure as for Example 1, 5.0 g) in isobutyric acid (20 mL) wasstirred at 120° C. overnight. The mixture was cooled to room temperatureand poured into water (100 mL). The pH was neutralized with aqueousNa₂CO₃ solution. It was then extracted with EtOAc and the organic layerwas washed with water and brine, dried over anhydrous MgSO₄, filteredand concentrated in-vacuo. The residue was then purified by silica gelchromatography eluted with EtOAc:petroleum ether=1:1 to afford thedesired product as a yellow solid (4.7 g, 77%). ¹H NMR (300 MHz, CDCl₃)δ ppm 1.51 (m, 6H, J=6.9 Hz), 3.32 (m, 1H, J=6.9 Hz), 8.16 (d, 1H, J=1.5Hz), 8.25 (d, 1H, J=1.5 Hz), 10.26 (br s, 1H); LC-MS: m/e=284 [M+1]+

b)6-bromo-2-(1-methylethyl)-1-(1-naphthalenylmethyl)-4-nitro-1H-benzimidazole

A mixture of 6-bromo-2-isopropyl-4-nitro-1H-benzo[d]imidazole (4.7 g),1-(bromomethyl)naphthalene (4.01 g) and K₂CO₃ (4.55 g) in DMF (150 mL)was stirred at 80° C. for 2 h. It was cooled to room temperature andfiltered. The filtrate was then poured into water (1 L). It was thenfiltered to afford a solid that was washed with water and then driedin-vacuo to afford the crude product (7.2 g). LC-MS: m/e=425 [M+1]+

c)2-(1-methylethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-nitro-1H-benzimidazole

A mixture of6-bromo-2-(1-methylethyl)-1-(1-naphthalenylmethyl)-4-nitro-1H-benzimidazole(2.05 g), morpholine (1.26 g), Pd₂(dba)₃ (0.46 g), Cs₂CO₃ (2.36 g) andX-Phos (0.41 g) in dioxane (30 mL) was degassed with nitrogen and thenstirred at 80° C. overnight. The mixture was cooled to room temperatureand the solvent was removed in-vacuo. The residue was then purified bysilica gel chromatography eluted with (EtOAc:petroleum ether=1:1) toafford the desired product as a yellow solid (1.6 g, 77%). LC-MS:m/e=431 [M+1]+

d)2-(1-methylethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-amine

TiCl₃ (16.3 mL) was added to a solution of2-(1-methylethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-nitro-1H-benzimidazole(1.6 g) and NH₄OAc (4 g) in MeOH (40 mL). After stirring for 4 h at roomtemperature, TLC showed no starting material. The pH of the mixture wasmade basic by adding Na₂CO₃ aqueous solution. The solvent was evaporatedoff under reduced pressure and the residue was extracted with DCM (250mL×2). The combined organic layers were washed with brine, dried overanhydrous Na₂SO₄, filtered and concentrated in-vacuo to afford thedesired product as a white solid (1.2 g, 81%). LC-MS: m/e=401 [M+1]+.

e)2-(1-methylethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-ol

To a solution of2-(1-methylethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-amine(300 mg) in H₂O (20 mL), and conc. H₂SO₄ (1 mL) was added aqueous NaNO₂(78 mg) solution dropwise at 0° C. The mixture was stirred at 0° C. for15 min and then heated to reflux for 1 h. The mixture was cooled to roomtemperature and the pH neutralized with aqueous NaHCO₃ solution. Thesolution was extracted with DCM (250 mL×3). The combined organic layerswere washed with brine, dried over anhydrous Na₂SO₄, filtered andconcentrated in-vacuo. The residue was then purified by silica gelchromatography eluted with EtOAc:petroleum ether=1:1 to afford thedesired product as a solid (80 mg, 27%). ¹H NMR (300 MHz, DMSO-d₆) δ ppm1.21 (d, 6H, J=6.6 Hz), 2.91 (t, 4H, J=7.5 Hz), 3.03 (m, 1H, J=6.6 Hz),3.64 (t, 4H, J=7.5 Hz), 5.89 (s, 2H), 6.27-6.34 (m, 3H), 7.32 (t, 1H,J=7.5 Hz), 7.59-7.70 (m, 2H), 7.83 (d, 1H, J=7.5 Hz), 8.00 (d, 1H, J=7.5Hz), 8.27 (d, 1H, J=7.5 Hz), 9.55 (s, 1H); LC-MS: m/e=402 [M+1]+.

Preparation of2-ethyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-ol

The titled compound was prepared following the same procedure as Example11 replacing isobutyric acid with propionic acid. ¹H NMR (300 MHz,DMSO-d₆) δ ppm 1.20 (t, 3H, J=7.5 Hz), 2.67 (q, 2H, J=7.5 Hz), 2.93 (t,4H, J=4.5 Hz), 3.64 (t, 4H, J=4.5 Hz), 5.86 (s, 2H), 6.26 (s, 1H),6.33-6.34 (m, 2H), 7.32 (t, 1H, J=7.5 Hz), 7.57-7.68 (m, 2H), 7.82 (d,1H, J=8.1 Hz), 7.98 (d, 1H, J=7.5 Hz), 8.23 (d, 1H, J=8.1 Hz), 9.54 (s,1H); LC-MS: m/e=388 [M+1]+.

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-(1-methylethyl)-6-(4-morpholinyl)-1H-benzimidazol-4-ol

The titled compound was prepared following the same procedure as Example11 replacing 1-(bromomethyl)naphthalene with1-(bromomethyl)-2,3-dichlorobenzene. ¹H NMR (300 MHz, CDCl₃) δ ppm 1.20(d, 6H, J=6.9 Hz), 2.94-3.06 (m, 2H), 3.67 (t, 4H, J=4.5 Hz), 5.45 (s,2H), 6.24-6.26 (m, 2H), 6.34 (s, 1H), 7.23 (d, 1H, J=7.8 Hz), 7.56 (d,1H, J=7.8 Hz), 9.54 (s, 1H); LC-MS: m/e=420 [M+1]+.

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-ethyl-6-(4-morpholinyl)-1H-benzimidazol-4-ol

The titled compound was prepared following the same procedure as Example12 replacing 1-(bromomethyl)naphthalene with1-(bromomethyl)-2,3-dichlorobenzene. ¹H NMR (300 MHz, CDCl₃) δ ppm 1.29(t, 3H, J=7.5 Hz), 2.86 (q, 2H, J=7.5 Hz), 3.10 (t, 4H, J=4.8 Hz), 3.82(t, 4H, J=4.8 Hz), 5.32 (s, 2H), 6.10 (d, 1H, J=2.4 Hz), 6.37 (dd, 2H,J=1.5, 7.8 Hz), 6.54 (d, 1H, J=2.4 Hz), 7.04 (t, 1H, J=7.8 Hz), 7.4 (dd,1H, J=1.5, 7.8 Hz); LC-MS: m/e=406 [M+1]+

Preparation of4-fluoro-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole

To a solution of2-methyl-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazol-4-amine(prepared following the same procedure used for Example 2) (200 mg) in70% HF/pyridine (2 mL) in a Teflon reactor was added NaNO₂ (56 mg) at−50° C. and the mixture was stirred at −50° C. for 30 min and thenheated to 70° C. for 1 h. The mixture was cooled to room temperature andthe pH neutralized with aqueous Na₂CO₃ solution. It was then extractedwith DCM (100 mL×2). The combined organic layers were concentratedin-vacuo and the residue was purified by Prep-TLC developed withEtOAc:petroleum ether=1:1 to afford the desired product as a solid (10mg, 5%). ¹H NMR (300 MHz, CDCl₃) δ ppm 2.54 (s, 3H), 3.05 (t, 4H, J=4.8Hz), 3.79 (t, 4H, J=4.8 Hz), 5.75 (s, 2H), 6.37 (d, 1H, J=1.8 Hz), 6.53(d, 1H, J=7.5 Hz), 6.66-6.71 (m, 1H), 7.30 (t, 1H, J=7.5 Hz), 7.59-7.69(m, 2H), 7.81 (d, 1H, J=9.0 Hz), 7.96 (d, 1H, J=7.5 Hz), 8.05 (d, 1H,J=7.8 Hz); LC-MS: m/e=376 [M+1]⁺.

Preparation of1-[(2,3-dichlorophenyl)methyl]-4-fluoro-2-methyl-6-(4-morpholinyl)-1H-benzimidazole

To a solution of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazol-4-amine(prepared following the same procedure as for Example 7, 200 mg) in 70%HF/pyridine (4 mL) in a Teflon reactor was added NaNO₂ (53 mg) at −50°C. and the mixture was stirred at −50° C. for 30 min and then heated to70° C. for 1 h. The mixture was cooled to room temperature and the pHwas neutralized with aqueous Na₂CO₃ solution. It was then extracted withDCM (100 mL×2). The combined organic layers were concentrated in-vacuoand the residue was purified by Prep-TLC developed with MeOH:DCM=1:30 toafford the desired product as a solid (40 mg, 20%). ¹H NMR (300 MHz,CDCl₃) δ 2.49 ppm (s, 3H), 3.09 (t, 4H, J=4.5 Hz), 3.83 (t, 4H, J=4.5Hz), 5.32 (s, 2H), 6.29-6.33 (m, 2H), 6.66 (d, 1H, J=7.8 Hz), 7.06 (t,1H, J=7.8 Hz), 7.42 (d, 1H, J=7.8 Hz); LC-MS: m/e=394 [M+1]⁺

Preparation of2-ethyl-4-fluoro-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole

To a solution of2-ethyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-amine(prepared following the same procedure as for Example 12, 200 mg, 0.49mmol) in 70% HF/pyridine (3 mL) was added NaNO₂ (50 mg, 0.73 mmol) at−50° C. and the resulting mixture was further stirred for 1 h. Then themixture was heated to 70° C. for 1 h. The mixture was cooled to rt andextracted with DCM (30 mL×3). The combined organic layers were driedover Na₂SO₄, concentrated in-vacuo. The resulting residue was purifiedby Prep-TLC to give the product (20 mg, 10%), as a white solid. ¹H NMR(300 MHz, DMSO-d₆) δ ppm 1.22 (t, 3H, J=7.5 Hz), 2.69 (q, 2H, J=7.5 Hz),3.02 (t, 4H, J=3.9 Hz), 3.66 (t, 4H, J=3.9 Hz), 5.97 (s, 2H), 6.32 (d,1H, J=7.8 Hz), 6.74-6.79 (m, 2H), 7.34 (t, 1H, J=7.8 Hz), 7.60-7.70 (m,2H), 7.85 (d, 1H, J=8.4 Hz), 8.00 (d, 1H, J=7.8 Hz), 8.24 (d, 1H, J=7.8Hz); LC-MS: m/e=390 [M+1]⁺.

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-ethyl-4-fluoro-6-(4-morpholinyl)-1H-benzimidazole

To a solution of1-[(2,3-dichlorophenyl)methyl]-2-ethyl-6-(4-morpholinyl)-1H-benzimidazol-4-amine(prepared following the same procedure as for Example 14, 203 mg) in 70%HF/pyridine (2 mL) in a Teflon reactor was added NaNO₂ (52 mg) at −50°C. and the mixture was stirred at −50° C. for 30 min and then heated to70° C. for 1 h. The mixture was cooled to room temperature and the pHneutralized with aqueous Na₂CO₃ solution. It was then extracted with DCM(100 mL×2). The combined organic layers were concentrated in-vacuo andthe residue was purified by Prep-TLC developed with EtOAc:peteroliumether=1:1 to afford the desired product as a solid (5 mg, 4%). ¹H NMR(300 MHz, CDCl₃): δ 1.39 ppm (t, 3H, J=7.5 Hz), 2.80 (q, 2H, J=7.5 Hz),3.10 (t, 4H, J=4.8 Hz), 3.84 (t, 4H, J=4.8 Hz), 5.34 (s, 2H), 6.29-6.34(m, 2H), 6.68 (dd, 1H, J=1.8, 12.6 Hz), 7.06 (t, 1H, J=7.8 Hz), 7.43 (d,1H, J=7.8 Hz); LC-MS: m/e=408 [M+1]⁺.

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-pyrazol-5-yl)-1H-benzimidazolea)4-bromo-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole

To a solution of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-amine(prepared following the same procedure as for Example 2, 1.1 g, 3 mmol)in aqueous HBr (50 mL) was added aqueous NaNO₂ (214 mg, 3.1 mmol)solution dropwise at 0-5° C. After addition the mixture was stirred at0° C. for 5 minutes, it was added to another solution of NaBr (927 mg, 9mmol) in aqueous HBr (50 mL) dropwise at 60° C. The resulting mixturewas then heated to 80° C. for 30 minutes and then cooled to roomtemperature. The solution pH was neutralized with aqueous NaHCO₃ (600mL) and extracted with DCM (500 mL×3). The combined organic layers wereconcentrated in vacuum and the residue was purified by silica gelchromatography eluted with petroleum ether:EtOAc=1:1 to give the desiredproduct (725 mg, 55%) as a white solid. ¹H NMR (300 MHz, CDCl₃) δ 2.55ppm (s, 3H), 3.05 (t, 4H, J=4.8 Hz), 3.79 (t, 4H, J=4.8 Hz), 5.73 (s,2H), 6.50 (dd, 1H, J=1.2, 7.5 Hz), 6.53 (d, 1H, J=1.8 Hz), 7.15 (d, 1H,J=1.8 Hz), 7.28 (t, 1H, J=7.5 Hz), 7.60-7.67 (m, 2H), 7.81 (d, 1H, J=8.4Hz), 7.96 (d, 1H, J=7.5 Hz), 8.06 (d, 1H, J=8.4 Hz); LC-MS: m/e=436[M+1]⁺.

b)2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-pyrazol-5-yl)-1H-benzimidazole

A mixture of4-bromo-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole(200 mg, 0.46 mmol), 1H-pyrazol-5-ylboronic acid (100 mg, 0.92 mmol),Pd(dba)₂ (40 mg, 0.046 mmol), Cs₂CO₃ (300 mg, 0.92 mmol) and P(t-Bu)₃(10 wt % in hexane, 20 mg, 0.092 mmol) in dioxane (20 mL) and water (10mL), was stirred at 100° C. for 18 h under a nitrogen atmosphere. Thereaction mixture was cooled and then concentrated. The resulting residuewas purified by silica gel chromatography eluted with EtOAc to give theproduct (140 mg 72%), as a white solid. ¹H NMR showed this compound isin a form of tautomeric mixture (major tautomer/minor tautomer=5/3) ¹HNMR of the major tautomer (300 MHz, DMSO-d₆) δ ppm 2.45 (s, 3H), 3.08(s, 4H), 3.71 (s, 4H), 6.00 (s, 2H), 6.37 (d, 1H, J=7.2 Hz), 6.96 (s,1H), 7.24-7.72 (m, 6H), 7.83-7.87 (m, 1H), 8.01 (d, 1H, J=7.2 Hz), 8.25(d, 1H, J=4.2 Hz), 13.24 (br s, 1H); LC-MS: m/e=424 [M+1]⁺

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid a) methyl2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylate

A mixture of intermediate4-bromo-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole,prepared following the same procedure as for Example 19 (400 mg, 0.92mmol), dppf (51 mg, 0.092 mmol), Pd(AcO)₂ (20.6 mg, 0.092 mmol) andtriethylamine (111 mg, 1.1 mmol) in methanol (50 mL), was degassed withCO(g). Then the reaction mixture was stirred at 60° C. for 18 h under aCO(g) atmosphere. The reaction mixture was cooled, concentrated. Theresulting residue was purified by silica gel chromatography eluted withEA to give the desired product (170 mg, 45%), as a white solid. ¹H NMR(300 MHz, DMSO-d₆) δ ppm 2.42 (s, 3H), 3.05 (t, 4H, J=4.8 Hz), 3.69 (t,4H, J=4.8 Hz), 3.90 (s, 3H), 6.02 (s, 2H), 6.28 (d, 1H, J=8.4 Hz),7.29-7.39 (m, 3H), 7.60-7.71 (m. 2H), 7.85 (d, 1H, J=8.4 Hz), 8.01 (d,1H, J=8.4 Hz), 8.24 (d, 1H, J=8.4 Hz); LC-MS: m/e=416 [M+1]⁺

b)2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylate(170 mg, 0.41 mmol) and LiOH (172 mg, 4.1 mmol) in THF (15 mL) and water(10 mL), was stirred at 50° C. for 1 h. Then the pH of the reactionmixture was neutralized with 1N aq. HCl. Then the mixture was stirred atrt for 1 h, filtered to give the desired product (150 mg, 91%), as awhite solid. ¹H NMR (300 MHz, DMSO-d6) δ ppm 2.51 (s, 3H), 3.07 (t, 4H,J=4.8 Hz), 3.70 (t, 4H, J=4.8 Hz), 6.09 (s, 2H), 6.38 (d, 1H, J=7.8 Hz),7.32-7.46 (m. 3H), 7.60-7.73 (m, 2H), 7.87 (d, 1H, J=7.8 Hz), 8.02 (d,1H, J=8.4 Hz), 8.23 (d, 1H, J=8.1 Hz); LC-MS: m/e=402 [M+1]⁺

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazolea)4-bromo-1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole

A solution of NaNO₂ (0.37 g, 5.4 mmol) in water (0.5 ml) was added to asolution of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazol-4-amine(prepared following the same procedure as for Example 7, 2.0 g, 5 mmol)in HBr (60 mL) at 0-5° C. and stirred for 15 min. The mixture was addeddropwise to a solution of NaBr (1.5 g, 15 mmol) in HBr (60 ml) at 60°C., and then heated to 80° C. for 30 min. The mixture was cooled to rtand poured into a Na₂CO₃ solution (200 ml). The mixture was extractedwith DCM (100 mL×3). The combined organic layers were dried over Na₂SO₄,filtered, concentrated. The residue was purified by silica gelchromatography eluted with EtOAc to give the product (1 g, 44%), as awhite solid. ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.39 (s, 3H), 3.06 (t, 4H,J=4.8 Hz), 3.70 (t, 4H, J=4.8 Hz), 5.53 (s, 2H), 6.31 (dd, 1H, J=1.2,7.8 Hz), 7.02 (d, 1H, J=2.1 Hz), 7.09 (d, 1H, J=2.1 Hz), 7.26 (t, 1H,J=7.8 Hz), 7.60 (dd, 1H, J=1.2 Hz, 7.8 Hz); LC-MS: m/e=455 [M+1]⁺.

b)1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carbonitrile

A mixture of4-bromo-1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole(1 g, 2.2 mmol), Pd(dba)₂ (161 mg, 0.22 mmol), dppf (244 mg, 0.44 mmol),Zn(CN)₂ (1030 mg, 8.8 mmol), water (1 mL), Fe(OAc)₂ (191 mg, 1.1 mmol)and Zn powder (429 mg, 6.6 mmol) in DMF (50 mL) was stirred at 100° C.under N₂ for 20 h. When TLC showed no starting material remaining, thereaction mixture was quenched with water and extracted with EtOAc (100mL×3). The organic layer was washed with brine, dried over MgSO₄,concentrated. The resulting residue was purified by silica gelchromatography eluted with EtOAc to give the product (400 mg, 45%), as awhite solid. ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.43 (s, 3H), 3.11 (t, 4H,J=4.8 Hz), 3.72 (t, 4H, J=4.8 Hz), 5.60 (s, 2H), 6.30 (dd, 1H, J=1.2,8.1 Hz), 7.25 (t, 1H, J=8.1 Hz), 7.35 (d, 1H, J=2.1 Hz), 7.42 (d, 1H,J=2.1 Hz), 7.60 (dd, 1H, J=1.2, 8.1 Hz); LC-MS: m/e=401 [M+1]⁺.

c)11-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide

A solution of KOH (78 mg, 1.4 mmol) in water (10 mL) was added dropwiseto solution of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carbonitrile(280 mg, 0.7 mmol) and 30% H₂O₂ (3 mL) in THF (10 mL) at rt. The mixturewas heated at 50° C. for 2 h. When TLC showed no starting material left,the pH of the mixture was acidified to pH ca. 5 and extracted with EtOAc(50 mL×3). The organic layer was washed with brine, dried over MgSO₄,concentrated. The resulting residue was purified by silica gelchromatography eluted with EtOAc to give the product (150 mg, 51%), as awhite solid. ¹H NMR (300 Mhz, DMSO-d₆) δ ppm 2.44 (s, 3H), 3.11 (t, 4H,J=4.8 Hz), 3.71 (t, 4H, J=4.8 Hz), 5.60 (s, 2H), 6.31 (d, 1H, J=8.1 Hz),7.16 (br s, 2H), 7.25 (t, 1H, J=8.1 Hz), 7.35 (d, 1H, J=1.8 Hz), 7.41(d, 1H, J=1.8 Hz), 7.60 (d, 1H, J=8.1 Hz); LC-MS: m/e=419 [M+1]⁺

d)1-[(2,3-dichlorophenyl)methyl]-N-[(1E)-(dimethylamino)methylidene]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide

A solution of combined batches of11-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide(220 mg, 0.52 mmol) in DMF-DMA (15 ml) was stirred at 130° C. for 2 h.When TLC showed no starting material remaining, the mixture was cooledto rt and the solvent was removed under reduced pressure to give thecrude product (JS211561-105A1, 220 mg, 89%), as a yellow solid. LC-MS:m/e=474 [M+1]⁺

e)1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazole

Hydrazine monohydrate (2 mL) was added to a solution of1-[(2,3-dichlorophenyl)methyl]-N-[(1E)-(dimethylamino)methylidene]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide(220 mg, 0.46 mmol) in acetic acid (5 mL) and stirred at 130° C. for 20min. The reaction mixture was cooled to rt and poured into saturatedNaHCO₃ solution (15 mL). The mixture was extracted with DCM (30 mL×3).The combined organic layers were dried over MgSO₄, filtered andconcentrated. The resulting residue was purified by silica gelchromatography eluted with DCM:MeOH=30:1 to give the desired product(110 mg, 53%), as a white solid. ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.50(s, 3H), 3.12 (t, 4H, J=4.8 Hz), 3.75 (t, 4H, J=4.8 Hz), 5.62 (s, 2H),6.36 (d, 1H, J=8.1 Hz), 7.21 (s, 1H), 7.27 (t, 1H, J=8.1 Hz), 7.54 (s,1H), 7.61 (d, 1H, J=8.1 Hz), 8.08 (s, 1H), 13.80 (s, 1H); LC-MS: m/e=443[M+1]⁺.

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid a) methyl1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate

A mixture of4-bromo-1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole(prepared following the same procedure as for Example 21, 150 mg, 0.33mmol), dppf (18 mg, 0.033 mmol), Pd(AcO)₂ (14.8 mg, 0.066 mmol) andtriethylamine (37 mg, 0.363 mmol) in methanol (30 mL), was degassed withCO(g). Then the reaction mixture was stirred at 60° C. for 4 h under aCO(g) atmosphere. The reaction mixture was cooled, concentrated. Theresulting residue was purified by silica gel chromatography eluted withpetroleum ether:EtOAc=1:1 to give the desired product (65 mg, 45%), as awhite solid. ¹H NMR (300 MHz, CDCl₃) δ 2.58 ppm (s, 3H), 3.16 (t, 4H,J=4.8 Hz), 3.86 (t, 4H, J=4.8 Hz), 4.07 (s, 3H), 5.38 (s, 2H), 6.24 (d,1H, J=7.8 Hz), 6.77 (d, 1H, J=2.4 Hz), 7.03 (t, 1H, J=7.8 Hz), 7.42 (d,1H, J=7.8 Hz), 7.67 (d, 1H, J=2.4 Hz); LC-MS: m/e=434 [M+1]⁺

b)1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate(65 mg, 0.15 mmol) and LiOH (19 mg, 0.5 mmol) in THF (5 mL) and water (5mL), was stirred overnight at rt. Then the pH of the reaction mixturewas neutralized with 1N aq. HCl. Then the mixture was stirred at rt for1 h, filtered to give the product (47 mg, 74%, with about 10%monochloride as the impurity) as a white solid. ¹H NMR (300 MHz,DMSO-d₆) δ ppm 2.53 (s, 3H), 3.12 (t, 4H, J=4.5 Hz), 3.73 (t, 4H, J=4.5Hz), 5.66 (s, 2H), 6.42 (d, 1H, J=7.8 Hz), 7.26 (t, 1H, J=7.8 Hz), 7.42(s, 1H), 7.48 (s, 1H), 7.62 (d, 1H, J=7.8 Hz); LC-MS: m/e=420 [M+1]⁺

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-4-(1H-pyrazol-5-yl)-1H-benzimidazole

A mixture of4-bromo-1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole(prepared following the same procedure as for Example 21, 250 mg, 0.55mmol), 1H-pyrazol-5-ylboronic acid (64 mg, 0.57 mmol), Pd(dba)₂ (32 mg,0.055 mmol), Cs₂CO₃ (358 mg, 1.1 mmol) and P(t-Bu)₃ (10 wt % in hexane,110 mg, 0.055 mmol) in dioxane (16 mL) and water (8 mL), was stirred at80° C. for 3 h under a nitrogen atmosphere. The reaction mixture wascooled and then concentrated. The resulting residue was purified bysilica gel chromatography eluted with petroleum ether:EtOAc=1:1 to givethe crude product (122 mg). The crude product was purified by Prep-HPLCto the pure product (72 mg, 30%), as a white solid. ¹H NMR showed thiscompound is in a form of tautomeric mixture (major tautomer/minortautomer=5/3) ¹H NMR of the major tautomer (300 MHz, DMSO-d₆) δ ppm 2.46(s, 3H), 3.12-3.14 (m, 4H), 3.73-3.76 (m, 4H), 5.57 (s, 2H), 6.36 (d,1H, J=7.8 Hz), 6.97 (s, 1H), 7.20-7.28 (m, 2H), 7.37 (s, 1H), 7.53-7.61(m, 2H), 13.17 (s, 1H); LC-MS: m/e=442 [M+1]⁺

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carbonitrile

A mixture of4-bromo-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole(prepared following the same procedure as for Example 19, 300 mg, 0.69mmol), Pd(PPh3)4 (80 mg, 0.069 mmol) and Zn(CN)₂ (162 mg, 1.38 mmol) inDMF (30 mL) was stirred at 80° C. under N₂ for 18 h. After cooling tort, the mixture was poured into water and filtered. The filter cake waspurified by silica gel chromatography eluted with petroleumether:EtOAc=1:1 to give the product (180 mg, 68%), as a white solid. ¹HNMR (300 MHz, DMSO-d₆): δ 2.44 (s, 3H), 3.07 (t, 4H, J=4.5 Hz), 3.68 (t,4H, J=4.5 Hz), 6.04 (s, 2H), 6.32 (d, 1H, J=7.2 Hz), 7.31-7.40 (m, 3H),7.60-7.71 (m, 2H), 7.86 (d, 1H, J=8.4 Hz), 8.01 (d, 1H, J=7.2 Hz), 8.22(d, 1H, J=8.4 Hz); LC-MS: m/e=383 [M+1]⁺

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazolea)2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide

A solution of KOH (45 mg, 0.8 mmol) in water (10 mL) was added dropwiseto solution of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carbonitrile(prepared using the same procedure as Example 24, 150 mg, 0.4 mmol) and30% H₂O₂ (3 ml) in THF (15 mL) at rt. The mixture was heated to 35° C.for 1 h. When TLC showed no starting material remaining, water (50 mL)was added, then it was filtered. The filter cake was purified by silicagel chromatography eluted with petroleum ether:EtOAc=1:2 to give theproduct (115 mg, 72%), as a white solid. ¹H NMR (300 MHz, DMSO-d₆): δppm 2.47 (s, 3H), 3.05 (t, 4H, J=4.8 Hz), 3.69 (t, 4H, J=4.8 Hz), 6.05(s, 2H), 6.34 (d, 1H, J=7.2 Hz), 7.25 (d, 1H, J=2.4 Hz), 7.34 (t, 1H,J=7.8 Hz), 7.53 (d, 1H, J=2.4 Hz), 7.63-7.70 (m, 2H), 7.77 (d, 1H, J=3.0Hz), 7.86 (d, 1H, J=8.1 Hz), 8.01 (d, 1H, J=7.2 Hz), 8.24 (d, 1H, J=8.1Hz), 9.24 (d, 1H, J=3.0 Hz); LC-MS: m/e=401 [M+1]⁺

b)N-[(1E)-(dimethylamino)methylidene]-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide

A solution of combined batches of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide(150 mg, 0.38 mmol) in DMF-DMA (10 mL) was stirred at 130° C. for 2 h.When TLC showed no starting material remaining, the mixture was cooledto rt and the solvent was removed under reduced pressure to give thecrude product (130 mg, 76%), as a yellow solid. ¹H NMR (300 MHz,DMSO-d₆): δ ppm 2.39 (s, 3H), 3.02 (t, 4H, J=4.5 Hz), 3.13 (s, 3H), 3.20(s, 3H), 3.69 (t, 4H, J=4.5 Hz), 5.99 (s, 2H), 6.30 (d, 1H, J=7.8 Hz),7.12 (d, 1H, J=2.4 Hz), 7.33 (t, 1H, J=7.8 Hz), 7.54 (d, 1H, J=2.4 Hz),7.60-7.71 (m, 2H), 7.85 (d, 1H, J=7.8 Hz), 8.01 (d, 1H, J=7.8 Hz), 8.25(d, 1H, J=7.8 Hz), 8.54 (s, 1H); LC-MS: m/e=456 [M+1]⁺

c)2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazole

Hydrazine hydrate (3 mL) was added to a solution ofN-[(1E)-(dimethylamino)methylidene]-2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide(130 mg, 0.29 mmol) in acetic acid (10 mL) and stirred at 130° C. for 30min. The reaction mixture was cooled to rt and poured into saturatedNa₂CO₃ solution (20 mL). A filtration was performed, and the filter cakewas purified by silica gel chromatography eluted with EtOAc to give theproduct (88 mg, 72%), as a white solid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm2.50 (s, 3H), 3.09 (s, 4H), 3.71 (s, 4H), 6.06 (s, 2H), 6.37 (d, 1H,J=7.8 Hz), 7.20 (s, 1H), 7.34 (t, 1H, J=7.8 Hz), 7.54 (s, 1H), 7.61-7.72(m, 2H), 7.86 (d, 1H, J=8.4 Hz), 8.02 (d, 1H, J=7.8 Hz), 8.09 (s, 1H),8.25 (d, 1H, J=8.4 Hz), 13.85 (s, 1H); LC-MS: m/e=425 [M+1]⁺

Preparation of methyl2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylatea) 3-amino-5-chloro-2-nitrobenzoic acid

Under nitrogen, to a solution of t-BuOK (156.8 g) and Cu(OAc)₂ (3.6 g)in DMF (1.2 L) was added a solution of 5-chloro-2-nitrobenzoic acid(40.0 g) and MeONH₂HCl (33.2 g) in DMF (300 mL) at 0° C. After 3 h thereaction was quenched by addition of H₂O (2.5 L) and acidified with 10%HCl solution to pH=1. The mixture was extracted with EA (2 L×2) and thecombined organic layers were then washed with brine, dried overanhydrous Na₂SO₄, filtered and concentrated in-vacuo to afford the crudeproduct as a yellow solid (43.2 g, yield 100%). ¹H NMR (300 MHz, CDCl₃):δ ppm 6.88 (s, 1H, J=2.4 Hz), 6.91 (d, 1H, J=2.4 Hz), 8.08 (br s, 2H);LC-MS: m/e=217 [M+1]⁺.

b) methyl 3-amino-5-chloro-2-nitrobenzoate

A mixture of 3-amino-5-chloro-2-nitrobenzoic acid (43.2 g) and HATU(2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate Methanaminium, commercially available) (76 g) inMeOH (81 mL), Et₃N (83 mL) and THF (300 mL) was stirred at roomtemperature for 3 h. When TLC showed no starting material, the solventwas removed in-vacuo and the residue was then diluted with EtOAc (2 L).It was then washed with brine (1 L×3) and dried over anhydrous Na₂SO₄,filtered and concentrated in-vacuo. The residue was then purified bysilica gel chromatography eluted with EtOAc:petroleum ether=1:8 toafford the desired product as a yellow solid (29.5 g, yield 64%). ¹H NMR(300 MHz, CDCl₃): δ ppm 3.90 (s, 3H, s), 5.85 (br s, 2H), 6.80 (d, 1H,J=2.4 Hz), 6.90 (d, 1H, J=2.4 Hz); LC-MS: m/e=231 [M+1]⁺.

c) methyl 3-amino-5-(4-morpholinyl)-2-nitrobenzoate

A mixture of combined batches of methyl 3-amino-5-chloro-2-nitrobenzoate(39 g), morpholine (29.5 g) and K₂CO₃ (47 g) was stirred in DMF (200 ml)at 110° C. for 5 h. The mixture was cooled to room temperature andpoured into water (1 L). It was extracted with EtOAc (500 mL×3). Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄, filtered and concentrated in-vacuo to afford the desired productas a yellow solid (22 g, yield 46%). ¹H NMR (300 MHz, CDCl₃): δ ppm 3.31(t, 4H, J=4.8 Hz), 3.82 (t, 4H, J=4.8 Hz), 3.89 (s, 3H), 6.03 (d, 1H,J=2.4 Hz), 6.34 (d, 1H, J=2.4 Hz); LC-MS: m/e=282 [M+1]⁺.

d) methyl 2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate

To a solution of methyl 3-amino-5-(4-morpholinyl)-2-nitrobenzoate (22 g)stirring at reflux in HOAc (400 mL) was added iron powder in portions(13 g). After the addition, the mixture was stirred at reflux for 5 h.It was cooled to room temperature and the solvent was removed in-vacuo.The residue was neutralized with aqueous Na₂CO₃ solution (1 L). It wasextracted with EtOAc (500 mL×3). The combined organic layers were thenconcentrated in-vacuo and the residue was purified by silica gelchromatography eluted with MeOH:DCM=1:30 to afford the desired productas a solid (16.6 g, yield 77%). ¹H NMR (300 MHz, CDCl₃): δ ppm 2.67 (s,3H), 3.17 (t, 4H, J=4.8 Hz), 3.90 (t, 4H, J=4.8 Hz), 3.98 (s, 3H), 7.44(d, 1H, J=1.8 Hz), 7.54 (d, 1H, J=1.8 Hz); LC-MS: m/e=276 [M+1]⁺.

e) methyl2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylate

A mixture of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate (4.125 g),1-(bromomethyl)naphthalene (5 g) and K₂CO₃ (6.2 g) was stirred at 80° C.for 3 h. When TLC showed no starting material remaining, the mixture wascooled to room temperature and then poured into water (500 mL). It wasextracted with EtOAc (500 mL×3) and the combined organic layers werewashed with brine (500 mL×3) and then concentrated in-vacuo. The residuewas purified by silica gel chromatography eluted with MeOH:DCM=1:100 toafford the desired product as a yellow solid (4.6 g, 74%). ¹H NMR (300MHz, DMSO-d₆): δ ppm 2.42 (s, 3H), 3.04 (t, 4H, J=4.8 Hz), 3.68 (t, 4H,J=4.8 Hz), 3.90 (s, 3H), 6.02 (s, 1H), 6.28 (d, 1H, J=7.5 Hz), 7.29 (d,1H, J=2.4 Hz), 7.32 (d, 1H, J=7.5 Hz), 7.39 (d, 1H, J=2.4), 7.60-7.71(m, 2H), 7.84 (d, 1H, J=8.4 Hz), 8.01 (d, 1H, J=7.5 Hz), 8.24 (d, 1H,J=7.5 Hz); LC-MS: m/e=416 [M+1]⁺.

Example 27

Preparation of11-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide

The titled compound was prepared from1-[2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carbonitrileusing the same procedure described in Example 21, step c. ¹H NMR (300Mhz, DMSO-d₆) δ ppm 2.44 (s, 3H), 3.11 (t, 4H, J=4.8 Hz), 3.71 (t, 4H,J=4.8 Hz), 5.60 (s, 2H), 6.31 (d, 1H, J=8.1 Hz), 7.16 (br s, 2H), 7.25(t, 1H, J=8.1 Hz), 7.35 (d, 1H, J=1.8 Hz), 7.41 (d, 1H, J=1.8 Hz), 7.60(d, 1H, J=8.1 Hz); LC-MS: m/e=419 [M+1]⁺

Preparation of methyl1-[(2-fluoro-3-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate

A mixture of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate prepared asdescribed in Example 26, step d (500 mg, 1.82 mmol), K₂CO₃ (502 mg, 3.64mmol) and 1-(bromomethyl)-2-fluoro-3-methylbenzene (389 mg, 1.91 mmol)in DMF (25 mL) was stirred at 80° C. for 18 h. The reaction mixture wascooled, poured into water (100 mL) and extracted with EtOAc (50 mL×3).The combined organic layers were dried over Na₂SO₄ and concentrated. Theresulting residue was purified by silica gel chromatography eluted withEtOAc:MeOH=100:1 to give the desired product (350 mg, 48%) as a redsolid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.24 (d, 3H, J=1.8 Hz), 2.47 (s,3H), 3.09 (t, 4H, J=4.8 Hz), 3.75 (t, 4H, J=4.8 Hz), 3.86 (s, 3H), 5.51(s, 2H), 6.62 (t, 1H, J=7.5 Hz), 7.00 (t, 1H, J=7.5 Hz), 7.22 (t, 1H,J=7.5 Hz), 7.32 (d, 1H, J=2.4 Hz), 7.36 (d, 1H, J=2.4 Hz); LC-MS:m/e=398 [M+1]⁺

Preparation of1-[(2-fluoro-3-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl1-[(2-fluoro-3-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 28 (240 mg, 0.6 mmol) and 2 N LiOH (1.8mL, 3.6 mmol) in THF (20 mL), was stirred at 45° C. for 16 h. Thesolution was filtered; the filter cake was then dissolved in water (20mL) and added into formic acid to adjust the pH of the solution to 3-4.Then a filtration was performed to provide the product (160 mg, 70%), asa white solid. ¹H NMR (300 MJz, DMSO-d₆): δ ppm 2.24 (d, 3H, J=1.8 Hz),2.50 (s, 3H), 3.12 (t, 4H, J=4.8 Hz), 3.75 (t, 4H, J=4.8 Hz), 5.57 (s,2H), 6.74 (t, 1H, J=7.5 Hz), 7.03 (t, 1H, J=7.5 Hz), 7.24 (t, 1H, J=7.5Hz), 7.40 (d, 1H, J=2.4 Hz), 7.42 (d, 1H, J=2.4 Hz); LC-MS: m/e=384[M+1]⁺

Preparation of methyl2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate

A solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate prepared asdescribed in Example 26, step d (500 mg, 1.8 mmol),1-(bromomethyl)-2-methyl-3-(trifluoromethyl)benzene (483 mg, 1.9 mmol)and K₂CO₃ (497 mg, 3.6 mmol) in DMF (50 mL) was stirred at 80° C. for 3h. The reaction mixture was cooled to rt and poured into water (50 mL),extracted with EtOAc (30 mL×3). The combined organic layers were washedwith brine, dried over Na₂SO₄ and concentrated. The resulting residuewas purified by silica gel chromatography eluted with DCM:MeOH=50:1 togive the crude product (230 mg, yield 29%), as a white solid. ¹H NMR(300 MHz, DMSO-d₆): δ ppm 2.39 (s, 3H), 2.54 (s, 3H), 3.08 (t, 4H, J=4.8Hz), 3.72 (t, 4H, J=4.8 Hz), 3.89 (s, 3H), 5.57 (s, 2H), 6.27 (d, 1H,J=7.5 Hz), 7.22 (t, 1H, J=7.5 Hz), 7.27 (d, 1H, J=2.4 Hz), 7.38 (d, 1H,J=2.4 Hz) 7.60 (d, 1H, J=7.5 Hz); LC-MS: m/e=448 [M+1]⁺

Preparation of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

An aqueous solution of 2 N LiOH (1.2 mL) was added to a solution ofmethyl2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 30 (180 mg, 0.4 mmol) in THF (10 mL)and stirred at 50° C. for 1 h. When TLC showed no starting materialremaining, the mixture was cooled to rt and THF was removed underreduced pressure. The pH of the mixture was acidified to pH 3. Thesuspension was filtered and the filtrate was collected, and washed withwater (10 mL) to give the product as a white solid (152 mg, yield 88%).¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.46 (s, 3H), 2.54 (s, 3H), 3.10 (t,4H, J=4.8 Hz), 3.73 (t, 4H, J=4.8 Hz), 5.63 (s, 2H), 6.37 (d, 1H, J=7.8Hz), 7.26 (t, 1H, J=7.8 Hz), 7.35 (d, 1H, J=2.4 Hz), 7.44 (d, 1H, J=2.4Hz), 7.62 (d, 1H, J=7.8 Hz); LC-MS: m/e=434 [M+1]⁺.

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carbonitrile

The titled compound was prepared from4-bromo-1-[(2,3-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazoleusing the same procedure as described in Example 21, step b. ¹H NMR (300MHz, DMSO-d₆) δ ppm 2.43 (s, 3H), 3.11 (t, 4H, J=4.8 Hz), 3.72 (t, 4H,J=4.8 Hz), 5.60 (s, 2H), 6.30 (dd, 1H, J=1.2, 8.1 Hz), 7.25 (t, 1H,J=8.1 Hz), 7.35 (d, 1H, J=2.1 Hz), 7.42 (d, 1H, J=2.1 Hz), 7.60 (dd, 1H,J=1.2, 8.1 Hz); LC-MS: m/e=401 [M+1]⁺.

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide

To a solution of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 20 (100 mg) in DCM (20 mL) wasadded a drop of DMF. The solution was then cooled to 0° C. and thenOxalyl chloride (64 mg) was then added. The mixture was stirred at roomtemperature for 30 min. The solvent was removed in-vacuo to afford awhite solid that used directly in the next step. To the solid dissolvedin dry DCM (20 mL) was bubbled in NH₃ at 0° C. for 5 min. The mixturewas then concentrated in-vacuo to afford the desired product as a whitesolid (79 mg, 79%). ¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.47 (s, 3H), 3.05(t, 4H, J=4.5 Hz), 3.69 (t, 4H, J=4.5 Hz), 6.05 (s, 2H), 6.35 (d, 1H,J=7.5 Hz), 7.23 (s, 1H), 7.34 (t, 1H, J=7.5 Hz), 7.53 (s, 1H), 7.60-7.74(m, 3H), 7.86 (d, 1H, J=7.5 Hz), 8.01 (d, 1H, J=9.0 Hz), 8.24 (d, 1H,J=9.0 Hz), 9.22 (s, 1H); LC-MS: m/e=401 [M+1]⁺.

Preparation of methyl2-methyl-6-(4-morpholinyl)-1-(8-quinolinylmethyl)-1H-benzimidazole-4-carboxylate

A mixture of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (500 mg, 1.82 mmol), K₂CO₃ (502 mg, 3.64mmol) and 5-(bromomethyl)quinoline (424 mg, 1.91 mmol) in DMF (25 mL)was stirred at 80° C. for 18 h. The reaction mixture was cooled, pouredinto water (100 mL) and extracted with EtOAc (50 mL×3). The combinedorganic layers were dried by Na₂SO₄ and concentrated. The resultingresidue was purified by silica gel chromatography eluted withEtOAc:MeOH=100:1 to give the crude product (350 mg, 46%), finally it waspurified by Prep-HPLC to give the product (180 mg, 24%) as a red solid.¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.52 (s, 3H), 3.03 (t, 4H, J=4.8 Hz),3.70 (t, 4H, J=4.8 Hz), 3.88 (s, 3H), 6.08 (s, 2H), 6.87 (d, 1H, J=7.5Hz), 7.31 (d, 1H, J=2.1 Hz), 7.36 (d, 1H, J=2.1 Hz), 7.48 (t, 1H, J=7.5Hz), 7.66 (dd, 1H, J=4.2, 8.4 Hz), 7.93 (d, 1H, J=7.5 Hz), 8.44 (dd, 1H,J=1.8, 8.4 Hz), 9.05 (dd, 1H, J=1.8, 4.2 Hz); LC-MS: m/e=417 [M+1]⁺

Example 35

Preparation of2-methyl-6-(4-morpholinyl)-1-(8-quinolinylmethyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl2-methyl-6-(4-morpholinyl)-1-(8-quinolinylmethyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 34 (300 mg, 0.72 mmol) and 2 N LiOH(2.2 mL, 4.3 mmol) in THF (10 mL), was stirred at 45° C. for 16 h. Itwas filtered and the filter cake was dissolved in water (20 mL) and thenadded into formic acid to adjust the pH of the solution to 3-4. Then afiltration was performed to give the product (200 mg, 69%), as a whitesolid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.61 (s, 3H), 3.06 (t, 4H, J=4.8Hz), 3.71 (t, 4H, J=4.8 Hz), 6.13 (s, 2H), 7.03 (d, 1H, J=7.5 Hz), 7.41(s, 2H), 7.51 (t, 1H, J=7.5 Hz), 7.66 (dd, 1H, J=4.2, 8.4 Hz), 7.95 (d,1H, J=7.5 Hz), 8.45 (dd, 1H, J=1.8, 8.4 Hz), 9.05 (dd, 1H, J=1.8, 4.2Hz); LC-MS: m/e=403 [M+1]⁺

Preparation of1-[(3,4-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid a) methyl1-[(3,4-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate prepared asdescribed in Example 26, step d (0.22 g, 0.799 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added in4-(chloromethyl)-1,2-dimethylbenzene (0.185 g, 1.199 mmol) and potassiumcarbonate (0.331 g, 2.397 mmol). The resulting reaction mixture wasstirred at 80° C. for 3 h. It was cooled to room temperature and pouredinto water (30 mL). The mixture was extracted with EtOAc (50 mL×3). Thecombined organic phases were washed with brine (50 mL) and concentrated.The crude material was subjected to normal phase purification (0˜40%EtOAc/Hexane) then (0˜1% MeOH/DCM) to give the product (0.24 g, 76%). MS(ES+) m/e 394.0 [M+H]⁺.

b)1-[(3,4-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a mixture of methyl1-[(3,4-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate(0.24 g, 0.61 mmol) in Tetrahydrofuran (THF) (10 mL) was added inlithium hydroxide (5.99 mL, 11.99 mmol). The reaction was stirred at 50°C. for 2 h. The reaction was cooled to room temperature. The organicsolvent was removed in-vacuo. The precipitate was collected byfiltration. Water (20 mL) was added in. The mixture was acidified with 1N HCl. The resulting solid was filtered and washed with water and driedto give the product (0.16 g, 66%). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.18(s, 6H), 2.58-2.75 (s, 3H), 3.06-3.22 (m, 4H), 3.71-3.82 (m, 4H), 5.55(s, 2H), 6.94 (m, 1H), 7.04 (s, 1H), 7.11 (d, 1H, J=7.83 Hz), 7.52 (m,2H). MS (ES+) m/e 380.2[M+H]⁺

Example 37

Preparation of2-methyl-6-(4-morpholinyl)-1-(2-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid

The titled compound was prepared following the same procedure as Example36 replacing 4-(chloromethyl)-1,2-dimethylbenzene with2-(bromomethyl)naphthalene in the first step. ¹H NMR (400 MHz, DMSO-d₆)δ ppm 2.73 (br. s., 3H), 3.11-3.21 (m, 4H), 3.70-3.78 (m, 4H), 5.82 (br.s., 2H), 7.40 (d, J=7.83 Hz, 1H), 7.48-7.63 (m, 4H), 7.72 (s, 1H),7.82-7.97 (m, 3H). MS (ES+) m/e 401.9 [M+H]⁺.

Preparation of1-[(3,4-dichlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

The titled compound was prepared following the same procedure as Example36 replacing 4-(chloromethyl)-1,2-dimethylbenzene with4-(bromomethyl)-1,2-dichlorobenzene in the first step. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 2.55 (s, 3H), 3.00-3.18 (m, 4H), 3.61-3.83 (m, 4H), 5.56(s, 2H), 7.02 (dd, J=8.34, 2.02 Hz, 1H), 7.44 (s, 2H), 7.50 (d, J=2.02Hz, 1H), 7.60 (d, 1H). MS (ES+) m/e 420.2 [M+H]⁺

Example 39

Preparation of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazolea)2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide

To the mixture of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 31 (0.6 g, 1.384 mmol) inDichloromethane (DCM) (60 mL) was added in oxalyl chloride (0.485 mL,5.54 mmol) and followed by the addition of ten drops of DMF. Thereaction mixture was stirred at rt for 10 minutes and concentrated togive the acid chloride. To a mixture of the crude acid chloride inTetrahydrofuran (THF) (60 mL) was bubbled with NH₃ gas. The reaction wasstirred at rt for 10 minutes. Brine (20 mL) and EtOAc (60 mL) was addedin and the aqueous phase was extracted with EtOAc (60 mL). The organicphase was combined and concentrated (0.59 g, 99%). The crude product wasused in the next step. MS (ES+) m/e 433.1 [M+H]⁺

b)2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-1H-benzimidazole

A mixture of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxamide(0.52 g, 1.202 mmol) in N,N-dimethylformamide dimethyl acetal (30 mL,224 mmol) was stirred at 105° C. for 2 hours and the reaction iscomplete. The reaction was concentrated under reduced pressure. To thecrude material was added Acetic Acid (30 mL) and hydrazine monohydrate(0.264 mL, 8.42 mmol). The reaction mixture was stirred at 100° C. for 1h and concentrated. The crude was purified using silica gel (0˜2%MeOH/DCM) to give the product (0.245 g, 42%). ¹H NMR (400 MHz, DMSO-d₆)δ ppm 2.48 (s, 3H), 2.56 (s, 3H), 3.10-3.15 (m, 4H), 3.67-3.79 (m, 4H),5.62 (s, 2H), 6.37 (d, J=7.83 Hz, 1H), 7.19 (d, J=2.02 Hz, 1H), 7.26 (t,J=7.83 Hz, 1H), 7.55 (d, J=1.77 Hz, 1H), 7.62 (d, J=7.58 Hz, 1H), 8.09(s, 1H), 13.83 (s, 1H). MS (ES+) m/e 457.1[M+H]⁺.

Preparation of2-methyl-4-(3-methyl-1H-1,2,4-triazol-5-yl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazolea)2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carbonitrile

To the mixture of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide,prepared as described in Example 33 (0.4 g, 0.999 mmol) inDichloromethane (DCM) (50 mL) was added in POCl₃ (0.931 mL, 9.99 mmol)and followed by the addition of ten drops of DMF. The reaction wasstirred at rt for 1 h. The mixture was quenched with aqueous sodiumbicarbonate solution. The aqueous phase was extracted with DCM (100 mL).The combined organic phase was washed with Brine then dried (MgSO₄),filtered and the solvent removed in-vacuo. The crude was purified onsilica (20˜50% EtOAc/Hexane) to give the product (0.246 g, 64%). ¹H NMR(400 MHz, DMSO-d₆) δ ppm 2.45 (s, 3H), 2.96-3.15 (m, 4H), 3.62-3.72 (m,4H), 6.05 (s, 2H), 6.32 (d, J=7.07 Hz, 1H), 7.29-7.44 (m, 3H), 7.57-7.74(m, 2H), 7.86 (d, J=8.08 Hz, 1H), 8.02 (d, J=7.83 Hz, 1H) 8.22 (d, 1H).MS (ES+) m/z 383.2 [M+H]⁺.

b)2-methyl-4-(3-methyl-1H-1,2,4-triazol-5-yl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole

To the suspension of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carbonitrile(120 mg, 0.314 mmol) in n-butanol (15 mL) was added in acetic hydrazide(232 mg, 3.14 mmol) and potassium carbonate (434 mg, 3.14 mmol). Thereaction was stirred at reflux temperature for 4 days. DCM (50 mL) andwater (50 mL) were added in. The organic phase was washed with Brine (50mL×3), dried (MgSO₄), and the solvent was removed. The crude waspurified by reverse phase purification to provide the desired product(36 mg, 25%). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.71 (br. s., 3H), 1.78(br. s., 3H), 2.21-2.37 (m, 4H), 2.90-3.08 (m, 4H), 5.20 (s, 2H), 5.74(d, J=7.33 Hz, 1H), 6.17 (br. s., 1H), 6.51 (t, J=7.71 Hz, 1H),6.75-6.95 (m, 3H), 7.04 (d, J=8.08 Hz, 1H), 7.17 (d, J=8.08 Hz, 1H),7.43 (d, 1H). MS (ES+) m/e 439.1[M+H]⁺.

Preparation of1-[2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-yl]ethanonea)N,2-dimethyl-N-(methyloxy)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide

To the suspension of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 20 (200 mg, 0.498 mmol) indichloromethane (DCM) (30 mL) was added in oxalyl chloride (0.218 mL,2.491 mmol) and followed by ten drops of DMF. The reaction mixture wasstirred at rt for 10 min. The reaction mixture was concentrated underreduced pressure to give the acid chloride. To the mixture of the acidchloride in dichloromethane (DCM) (30 mL) was added inN,O-dimethylhydroxylamine hydrochloride (97 mg, 0.996 mmol) and TEA(0.694 mL, 4.98 mmol). The reaction mixture was stirred at rt for 18 h.Water (50 mL) was added in and the aqueous phase was extracted with DCM(50 mL×2). The combined organic phase was washed with Brine (50 mL),dried (MgSO₄) and concentrated under reduced pressure. The crude waspurified on a silica column purification (0˜4% MeOH/DCM) to give theproduct (100 mg, 43%). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.39 (s, 3H),2.92-3.10 (m, 4H), 3.26 (br. s., 3H), 3.56-3.75 (m, 7H), 5.99 (s, 2H),6.35 (d, J=6.57 Hz, 1H) 6.86 (d, J=2.02 Hz, 1H), 7.04 (d, J=1.77 Hz,1H), 7.26-7.40 (m, 1H), 7.54-7.73 (m, 2H), 7.86 (d, J=8.34 Hz, 1H), 8.02(d, J=7.07 Hz, 1H), 8.25 (d, 1H). MS (ES+) m/e 445.2[M+H]⁺.

b)1-[2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-yl]ethanone

To a solution ofN,2-dimethyl-N-(methyloxy)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide(82 mg, 0.184 mmol) in Tetrahydrofuran (THF) (10 mL) was added a 3.0 Msolution of methylmagnesium chloride (0.123 mL, 0.369 mmol) in THF at 0°C. The resulting reaction mixture was stirred at 0° C. for 2 h and thenquenched very carefully with saturated aqueous ammonium chloride. Themixture was diluted with ethyl acetate (50 mL) and the aqueous phase wasextracted with ethyl acetate (50 mL×2). The combined organic phases werewashed with brine (50 mL), dried over (MgSO₄), and filtered. Thesolution was concentrated under reduced pressure. The crude product waspurified on a silica column (40˜60% EtOAc/Hexane) to give the product(46 mg, 59%). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.47 (s, 3H), 3.00 (s,3H), 3.02-3.10 (m, 4H), 3.59-3.73 (m, 4H), 6.06 (s, 2H), 6.33 (d, J=6.82Hz, 1H), 7.21-7.42 (m, 3H), 7.52-7.76 (m, 2H), 7.86 (d, J=8.34 Hz, 1H),8.02 (d, J=7.33 Hz, 1H), 8.25 (d, 1H). MS (ES+) m/e 399.9[M+H]⁺.

Preparation of[2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazol-4-yl]methanol

To the mixture of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 20 (70 mg, 0.174 mmol) inTetrahydrofuran (THF) (5 mL) was added in LiAlH₄ (19.85 mg, 0.523 mmol)at 0° C. and the reaction mixture was stirred at rt for 1 h. Then LiAlH₄(19.85 mg, 0.523 mmol) was added in and the reaction mixture was stirredat rt for another hour. The reaction mixture was cooled to 0° C. andquenched with water (0.04 ml), NaOH (15%, 0.04 ml) then water (0.12 ml).After the resultant mixture was stirred at room temperature for 2 h,anhydrous MgSO₄ was added and the reaction mixture was filtered throughcelite and washed with EtOAc. Evaporation of the solvent gave the crudeproduct. The crude product was purified on a silica column (0-4%MeOH/DCM) to give the solid (12 mg, 17%). ¹H NMR (400 MHz, DMSO-d₆) δppm 2.38 (s, 3H), 2.91-3.05 (m, 4H), 3.64-3.74 (m, 4H), 4.89 (d, J=5.56Hz, 2H), 5.12 (t, J=5.81 Hz, 1H), 5.95 (s, 2H), 6.33 (d, J=6.82 Hz, 1H),6.81 (d, J=2.02 Hz, 1H), 6.97 (d, J=1.77 Hz, 1H), 7.34 (t, J=7.71 Hz,1H), 7.54-7.74 (m, 2H), 7.85 (d, J=8.08 Hz, 1H), 8.01 (d, J=7.58 Hz,1H), 8.25 (d, 1H). MS (ES+) m/z 388.0 [M+H]⁺.

Preparation of[2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazol-4-yl]methanol

A solution of methyl2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 30 (0.37 g, 0.827 mmol) inTetrahydrofuran (THF) (10 mL) was cooled to 0° C. LiAlH₄ (0.038 g, 1mmol) in THF (3 mL) was added in and the reaction mixture was stirred at0° C. for 30 min. The reaction mixture was quenched with water (0.04mL), NaOH (15%, 0.04 mL) then water (0.12 mL). Anhydrous MgSO₄ was addedand the reaction mixture was filtered through celite and washed withEtOAc. Evaporation of the solvent gave the crude product. The crudeproduct was purified on a silica column (1˜4% MeOH/DCM) to give thesolid (0.32 g, 88%). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.35 (s, 3H), 2.56(s, 3H), 2.99-3.07 (m, 4H), 3.67-3.76 (m, 4H), 4.87 (d, J=5.81 Hz, 2H),5.11 (t, J=5.68 Hz, 1H), 5.51 (s, 2H), 6.30 (d, J=7.83 Hz, 1H), 6.81 (d,J=2.27 Hz, 1H), 6.97 (d, J=2.02 Hz, 1H), 7.24 (t, J=7.83 Hz, 1H), 7.60(d, 1H). MS (ES+) m/z 420.1 [M+H]⁺.

Example 44

Preparation of2-methyl-N-(methylsulfonyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxamide

To the mixture of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 20 (100 mg, 0.249 mmol) inN,N-Dimethylformamide (DMF) (2 mL) in a 20 mL vial was added EDC (57.3mg, 0.299 mmol), methane sulfonamide (47.4 mg, 0.498 mmol) and DMAP(21.30 mg, 0.174 mmol). The mixture was stirred at 60° C. and monitoredby LC/MS. After stirring for 5 days the DMF was removed in-vacuo and theremaining residue was dissolved in 2 mL of DMSO and purified by reversephase chromatography with 2 injections eluting with a 27% to 57%AcCN/H₂O gradient over 12 minutes. The fractions containing the desiredcompound, as determined by LC/MS, were combined and concentratedin-vacuo to provide the desired compound (47 mg, 0.097 mmol, 39.0%yield) as bright yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.52 (s,3H), 3.12 (t, 4H), 3.50 (s, 3H), 3.73 (t, 4H), 6.15 (s, 2H), 6.38 (d,1H), 7.35 (t, aH), 7.50 (s. 1H), 7.51-7.75 (m, 2H), 7.88 (d, 1H), 8.05(d, 1H), 8.25 (2, 1H), 8.35 (s, 1H), 12.8 (br. s, 1H); LC-MS: m/e=480[M+1]⁺

Preparation of methyl5-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-7-carboxylate a)methyl 2,3-diamino-5-(4-morpholinyl)benzoate

A mixture of methyl 3-amino-5-morpholino-2-nitrobenzoate, prepared asdescribed in Example 26, step c (19.2 g, 68.3 mmol) and Pd/C (1.9 g) inMeOH (500 mL) in an autoclave under an atmosphere of H₂ (4 atm) werestirred at room temperature for 3 h, When TLC analysis indicatedcomplete consumption of starting material, the mixture was filtered andthe filtrate was concentrated in-vacuo to afford the desired product asa brown solid (13.9 g, 80.1%). ¹H NMR (300 MHz, DMSO-d₆): δ ppm 6.62 (d,1H, J=2.4 Hz), 6.54 (d, 1H, J=2.4 Hz), 5.84 (s, 2H), 4.75 (s, 2H), 3.76(s, 3H), 3.70 (t, 4H, J=4.8 Hz), 2.85 (t, 4H, 4.8 Hz). LC-MS: m/e=252.1[M+1]⁺.

b) methyl5-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-7-carboxylate

A mixture of methyl 2,3-diamino-5-(4-morpholinyl)benzoate (4.0 g) inCF₃COOH (20 mL) was heated at reflux temperature for 8 h. When TLCanalysis indicated consumption of starting material, the mixture wascooled to room temperature and the solvent was removed in-vacuo. Theresidue was diluted with aqueous NaHCO₃ and extracted with EtOAc (250mL×3). The combined organic layers were washed with brine (250 mL×2),dried over anhydrous Na₂SO₄. After filtration, the solvent was removedby rotary evaporator. The residue was then purified by silica gelchromatography eluted with EtOAc:petroleum ether=1:4 to afford thedesired product as a pale solid. (4.3 g, 82.7%). ¹H NMR (300 MHz,DMSO-d₆): δ ppm 13.47 (s, 1H), 7.71 (s, 1H), 7.56 (s, 1H), 3.96 (s, 3H),3.79 (t, 4H, J=4.5 Hz), 3.17 (s, 4H). LC-MS: m/e=330.1 [M+1]⁺.

Example 46

Preparation of methyl1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate

A suspension of methyl5-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-7-carboxylate,prepared as described in Example 45 (1.5 g, 4.56 mmol) and potassiumcarbonate (1.889 g, 13.67 mmol) in N,N-Dimethylformamide (DMF) (10 mL)was stirred at rt for 15 min.1-(Bromomethyl)-2-methyl-3-(trifluoromethyl)benzene (1.729 g, 6.83 mmol)was added in and the resulting reaction mixture was stirred for 3 h at80° C. The mixture was then cooled to room temperature and poured intoice/water. The precipitate was collected by filtration, washed withwater, then hexanes (turned into a gum on the filter paper—some materialwas lost). The crude material was purified on a silica gel column (ISCO,eluting with 0-5% MeOH in DCM) to give the desired product (580 mg,1.099 mmol, 24.12% yield) (several mixed fractions obtained werediscarded). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.65 (d, J=2.53 Hz, 1H),7.61 (d, J=7.83 Hz, 1H), 7.37 (d, J=2.27 Hz, 1H), 7.23 (t, J=7.96 Hz,1H), 6.29 (d, J=7.58 Hz, 1H), 5.76 (s, 2H), 3.93 (s, 3H), 3.70-3.78 (m,4H), 3.12-3.22 (m, 4H), 2.53 (s, 3H). MS (ES+) m/e 502 [M+H]⁺.

Preparation of1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 46 (510 mg, 1.017 mmol) and 2 M lithiumhydroxide (6 mL, 12.00 mmol) in THF (12 mL) was stirred at 50° C. for 2h. The reaction was cooled to room temperature. The organic solvent wasremoved under reduced pressure and the aqueous was diluted with waterand acidified by the addition of 1 N HCl. The precipitate formed wascollected by filtration. The solid was washed with ether and turned intoa gummy residue. The residue was washed with MeOH until all the materialwas transferred into the collection flask. The organics were evaporatedand a white solid formed upon standing. The precipitate was collected byfiltration, washed with water and dried to give the desired product (438mg, 0.881 mmol, 87% yield) as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ ppm 7.68 (d, J=2.02 Hz, 1H), 7.61 (d, J=8.08 Hz, 1H), 7.23 (t, J=7.83Hz, 1H), 7.08 (d, J=2.02 Hz, 1H), 6.27 (d, J=7.58 Hz, 1H), 5.74 (s, 2H),3.63-3.82 (m, 4H), 3.05-3.21 (m, 4H), 2.52 (br. s., 3H). MS (ES+) m/e488 [M+H]⁺.

Preparation of methyl6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate

A mixture of methyl5-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-7-carboxylateprepared as described in Example 45 (1.5 g, 4.56 mmol) and potassiumcarbonate (1.889 g, 13.67 mmol) in N,N-Dimethylformamide (DMF) (10 mL)was stirred at rt for 10 min. After addition of1-(bromomethyl)naphthalene (1.511 g, 6.83 mmol), the mixture was warmedto 80° C. and stirred for 3 h at this temperature. The resulting mixturewas cooled to rt and poured over ice. The precipitate formed wascollected by filtration and air dried (2.4 g total). The crude materialwas purified on silica gel (ISCO, 0-5% MeOH in DCM) to give methyl6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate(1.48 g, 3.15 mmol, 69.2% yield). A portion of this material (138 mg)was purified by reverse phase-HPLC (25 to 95% AcCN in water, plus 0.1%TFA) to give the desired product (93.4 mg, 0.195 mmol, 4.28% yield) as awhite solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.26 (d, J=8.34 Hz, 1H),8.01 (d, J=7.33 Hz, 1H), 7.85 (d, J=8.08 Hz, 1H), 7.60-7.73 (m, 3H),7.39 (d, J=2.27 Hz, 1H), 7.31 (t, J=7.83 Hz, 1H), 6.24 (br. s., 1H),6.22 (s, 2H), 3.95 (s, 3H), 3.64-3.73 (m, 4H), 3.06-3.19 (m, 4H). MS(ES+) m/e 470 [M+H]⁺.

Preparation of methyl1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate

A mixture of methyl5-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-7-carboxylate,prepared as described in Example 45 (1.5 g, 4.56 mmol) and potassiumcarbonate (1.889 g, 13.67 mmol) in N,N-Dimethylformamide (DMF) (10 mL)was stirred at rt for 10 min. After addition of1-(bromomethyl)-3-chloro-2-methylbenzene (1.500 g, 6.83 mmol), themixture was warmed to 80° C. and stirred for 3 h at this temperature.The resulting mixture was cooled to rt and poured over ice. Theprecipitate formed was collected by filtration and air dried (2.4 gtotal). Purification on a silica gel column (10-50% EtOAc in hexane)failed to produce pure material. The fractions containing product werecombined and the solvent was removed under reduced pressure to affordmethyl1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate(2.03 g, 4.34 mmol, 95% yield) (only 87% pure). A portion of thismaterial (165 mg) was purified by RP-HPLC (25 to 95% AcCN in water, plus0.1% TFA) to give pure desired product (92.3 mg, 0.193 mmol, 4.24%yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.64 (d, J=2.27 Hz, 1H),7.28-7.43 (m, 2H), 7.04 (t, J=7.96 Hz, 1H), 5.97 (d, J=7.83 Hz, 1H),5.71 (s, 2H), 3.93 (s, 3H), 3.66-3.80 (m, 4H), 3.06-3.25 (m, 4H), 2.46(s, 3H). MS (ES+) m/e 468 [M+H]⁺

Preparation of6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylicacid

A suspension of methyl6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 48 (1.28 g, 2.73 mmol) in Methanol (18mL) and 1 M sodium hydroxide (15 mL, 15.00 mmol) was stirred overnightat rt, then at 50° C. for 5 h. The reaction was cooled to roomtemperature and acidified (pH 4) by the addition of 1 N HCl. Theprecipitate formed was collected by filtration, washed with water anddried to give the desired product (1.13 g, 2.233 mmol, 82% yield). Aportion of this material (132 mg) was purified by reverse phase-HPLC(15-95% AcCN in water plus 0.1% TFA). The fractions containing productwere combined and the volume was reduced to about ⅓ of the original. Theprecipitate formed was collected, washed with water and dried in avacuum oven (50° C., overnight) to give the desired product (88.4 mg,0.194 mmol, 7.12% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.11 (br. s.,1H), 8.26 (d, J=8.59 Hz, 1H), 8.01 (d, J=7.33 Hz, 1H), 7.85 (d, J=8.34Hz, 1H), 7.57-7.73 (m, 3H), 7.35 (d, J=2.53 Hz, 1H), 7.29-7.34 (m, 1H),6.24 (d, J=7.07 Hz, 1H), 6.22 (s, 2H), 3.64-3.72 (m, 4H), 3.06-3.18 (m,4H). MS (ES+) m/e 456 [M+H]⁺.

Preparation of methyl1-[(2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate

A mixture of methyl5-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-7-carboxylate,prepared as described in Example 45 (1.5 g, 4.56 mmol) and potassiumcarbonate (1.889 g, 13.67 mmol) in N,N-Dimethylformamide (DMF) (10 mL)was stirred at rt for 10 min. After addition of1-(bromomethyl)-2,3-dichlorobenzene (1.639 g, 6.83 mmol), the mixturewas warmed to 80° C. and stirred for 3 h at this temperature. Theresulting mixture was cooled to rt and poured over ice. The precipitateformed was collected by filtration and air dried to give crude product(2.2 g, 4.51 mmol, 99% yield) (91% pure). A portion of this material(230 mg) was purified by reverse phase-HPLC (25 to 95% AcCN in water,plus 0.1% TFA) to give the desired product (137.4 mg, 0.276 mmol, 6.05%yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.65 (d, J=2.27 Hz, 1H), 7.61(dd, J=8.08, 1.26 Hz, 1H), 7.48 (d, J=2.27 Hz, 1H), 7.24 (t, J=7.96 Hz,1H), 6.25 (dd, J=7.83, 1.26 Hz, 1H), 5.77 (s, 2H), 3.93 (s, 3H),3.68-3.81 (m, 4H), 3.13-3.24 (m, 4H). MS (ES+) m/e 488 [M+H]⁺.

Preparation of1-[(2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl1-[(2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylateprepared as described in Example 51 (1.95 g, 3.99 mmol) and 2 M lithiumhydroxide (0.096 g, 3.99 mmol) in Tetrahydrofuran (THF) was stirred at50° C. for 3 h. The reaction was cooled to room temperature, the organicsolvent was removed under reduced pressure and the aqueous residue wasacidified (pH 4) by the addition of 1N HCl. A gummy precipitate formed.After standing at rt overnight, it turned into a solid. The precipitatewas collected, washed with water and dried to give crude desired product(1.83 g, 3.86 mmol, 97% yield) as a gray solid. A portion of thismaterial (148 mg) was purified by reverse phase-HPLC (15-95% AcCN inwater plus 0.1% TFA). The fractions containing product were combined andthe volume was reduced to about ⅓ of the original. The precipitateformed was collected, washed with water and dried in a vacuum oven togive desired product (88.3 mg, 0.182 mmol, 4.57% yield). ¹H NMR (400MHz, DMSO-d₆) δ ppm 13.09 (s, 1H), 7.64 (d, J=2.53 Hz, 1H), 7.61 (dd,J=8.08, 1.26 Hz, 1H), 7.44 (d, J=2.27 Hz, 1H), 7.24 (t, J=8.08 Hz, 1H),6.26 (dd, J=7.83, 1.26 Hz, 1H), 5.77 (s, 2H), 3.66-3.81 (m, 4H),3.12-3.25 (m, 4H). MS (ES+) m/e 474 [M+H]⁺.

Preparation of1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylicacid

A suspension of methyl1-[3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 49 (1.09 g, 2.330 mmol) in Methanol (12mL) and Tetrahydrofuran (THF) (4 mL) was treated with 1 M aq. sodiumhydroxide (12 mL, 12.00 mmol) and stirred at 70° C. for 1.5 h (mixtureturned homogeneous). The reaction was cooled to room temperature, thevolume reduced to half and the residue was acidified (pH 4) by theaddition of 1 N HCl. The precipitate was collected, washed with waterand dried to give crude desired product (918.6 mg, 2.024 mmol, 87%yield) as a yellow solid. A portion of it (140 mg) was suspended in 3.5mL of DMSO. After sonication and heating, the solid went into solutionbut it crashed out. The precipitate was collected and washed with DMSO,but it still showed impurities by LC/MS. Another aliquot (132 mg) wasdissolved with heating in 5 mL of DMSO and purified by reversephase-HPLC (15 to 95% AcCN in water plus 0.1% TFA). The desired productwas isolated by evaporation of the organic solvent: the precipitate wascollected by filtration, washed with water and dried in a vacuum oven toafford pure desired product (89 mg, 0.192 mmol, 8.25% yield). ¹H NMR(400 MHz, DMSO-d₆) δ ppm 13.09 (s, 1H), 7.63 (d, J=2.27 Hz, 1H), 7.36(d, J=7.83 Hz, 1H), 7.30 (d, J=2.27 Hz, 1H), 7.05 (t, J=7.96 Hz, 1H),5.98 (d, J=7.58 Hz, 1H), 5.70 (s, 2H), 3.66-3.83 (m, 4H), 3.08-3.24 (m,4H), 2.47 (s, 3H). MS (ES+) m/e 453.9 [M+H]⁺.

Preparation of1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxamide

Oxalyl chloride (0.251 mL, 2.87 mmol) was added to a suspension of1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 47 (350 mg, 0.718 mmol) inDichloromethane (DCM) (6 mL). The reaction mixture was stirred at rt for10 minutes (turned into a solution) and then the solvent was evaporated.The residue (crude acid chloride), was dissolved in Tetrahydrofuran(THF) (6 mL). NH₃ gas was bubbled in (the mixture changed color yellowto white, and a precipitate formed); the mixture was stirred at rt for10 minutes and then partitioned between brine (15 mL) and EtOAc (20 mL).The aqueous phase was extracted with another aliquot of EtOAc (20 mL).The organic phases were combined and concentrated to afford desiredproduct (297 mg, 0.580 mmol, 81% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm8.59 (d, J=2.53 Hz, 1H), 7.99 (d, J=2.78 Hz, 1H), 7.77 (d, J=2.27 Hz,1H), 7.62 (d, J=7.58 Hz, 1H), 7.31 (d, J=2.27 Hz, 1H), 7.25 (t, J=7.83Hz, 1H), 6.36 (d, J=7.58 Hz, 1H), 5.78 (s, 2H), 3.60-3.80 (m, 4H),3.09-3.22 (m, 4H), 2.53 (s, 3H). MS (ES+) m/e 487 [M+H]⁺

Example 55

Preparation of1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-2-(trifluoromethyl)-1H-benzimidazole

A mixture of1-{[2-Methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxamide,prepared as described in Example 54 (250 mg, 0.514 mmol) andN,N-dimethylformamide dimethyl acetal (7 mL, 52.3 mmol) was stirred at105° C. for 1.5 hour. The reaction mixture was concentrated underreduced pressure and the residue was suspended in Acetic Acid (5 mL).After the addition of hydrazine monohydrate (0.113 mL, 3.60 mmol) thereaction mixture was heated at 100° C. for 1 hour. The solvent wasconcentrated under vacuum, the residue was azeotroped with toluene (2×),and the residue was dissolved in DMSO and purified by reverse phase-HPLC(20-95% AcCN in water plus 0.1% TFA). The fractions containing productwere combined, neutralized by the addition of aq NaHCO₃ sat sol and theorganic evaporated. The precipitate in the aqueous residue was collectedby filtration, washed with water and dried in a vacuum oven (45° C.)overnight to give the desired product (157.1 mg, 0.302 mmol, 58.7%yield) as a white powder. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 13.05(br. s., 1H), 8.12 (s, 1H), 8.00 (d, J=2.27 Hz, 1H), 7.63 (d, J=7.83 Hz,1H), 7.18 (t, J=7.83 Hz, 1H), 6.56 (d, J=7.83 Hz, 1H), 6.51 (d, J=2.27Hz, 1H), 5.54 (s, 2H), 3.71-3.95 (m, 4H), 3.09-3.30 (m, 4H), 2.57 (s,3H). MS (ES+) m/e 511[M+H]⁺.

Example 56

Preparation of1-[(2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-2-(trifluoromethyl)-1H-benzimidazolea)1-[(2,3-Dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxamide

Oxalyl chloride (0.332 mL, 3.80 mmol) was added to a suspension of1-[(2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 52 (450 mg, 0.949 mmol) inDichloromethane (DCM) (7 mL). The reaction mixture was stirred at rt for10 minutes and then the solvent was evaporated. The residue (crude acidchloride), was suspended in Tetrahydrofuran (THF) (7 mL). NH₃ gas wasbubbled in (the mixture changed color yellow to white), the mixture wasstirred at rt for 15 minutes and then partitioned between brine (15 mL)and EtOAc (20 mL). The aqueous phase was extracted with EtOAc (2×20 mL)and CH₂Cl₂ (10 mL). The organic phases were combined, dried over Na₂SO₄and concentrated to afford the desired product (395 mg, 0.835 mmol, 88%yield) which was used as is in the next step. ¹H NMR (400 MHz, DMSO-d₆)δ ppm 8.56 (d, J=2.27 Hz, 1H), 7.98 (d, J=2.53 Hz, 1H), 7.76 (d, J=2.27Hz, 1H), 7.62 (dd, J=8.08, 1.26 Hz, 1H), 7.42 (d, J=2.27 Hz, 1H), 7.25(t, J=7.96 Hz, 1H), 6.33 (dd, J=7.83, 1.26 Hz, 1H), 5.80 (s, 2H),3.68-3.83 (m, 4H), 3.14-3.24 (m, 4H). MS (ES+) m/e 473.1 [M+H]⁺.

b)1-[(2,3-Dichlorophenyl)methyl]-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-2-(trifluoromethyl)-1H-benzimidazole

A suspension of1-[(2,3-dichlorophenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxamide,prepared as described in Example 56, step a (389 mg, 0.822 mmol) inN,N-dimethylformamide dimethyl acetal (9 mL, 67.2 mmol) was stirred at105° C. for 1 hour. The reaction was concentrated under reduced pressureand the residue was suspended in Acetic Acid (7 mL). After the additionof hydrazine monohydrate (0.181 mL, 5.75 mmol) the reaction mixture washeated at 100° C. for 1 hour. The solvent was concentrated under vacuumand the residue was dissolved in DMSO and purified by reverse phase-HPLC(20-95% AcCN in water plus 0.1% TFA) (some residual solid filtered off).The fractions containing product were combined, neutralized by theaddition of aq NaHCO₃ sat sol and the organic evaporated (some compoundlost during transfer). The precipitate in the aqueous residue wascollected by filtration, washed with water and dried in a vacuum oven(45° C.) overnight to give the desired product (115 mg, 0.227 mmol,27.6% yield) as a white powder. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm13.05 (br. s., 1H), 8.12 (s, 1H), 8.00 (d, J=2.27 Hz, 1H), 7.47 (dd,J=8.08, 1.26 Hz, 1H), 7.10 (t, J=7.96 Hz, 1H), 6.59 (d, J=2.27 Hz, 1H),6.37 (dd, J=7.83, 1.01 Hz, 1H), 5.62 (s, 2H), 3.79-3.92 (m, 4H),3.17-3.30 (m, 4H). MS (ES+) m/e 497 [M+H]⁺.

Preparation of1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-4-(1H-1,2,4-triazol-3-yl)-2-(trifluoromethyl)-1H-benzimidazole

Oxalyl chloride (0.347 mL, 3.97 mmol) was added to a suspension of1-[(3-chloro-2-methylphenyl)methyl]-6-(4-morpholinyl)-2-(trifluoromethyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 53 (450 mg, 0.992 mmol) inDichloromethane (DCM) (8 mL). The reaction mixture was stirred at rt for10 minutes and then the solvent was evaporated. The residue (crude acidchloride), was suspended in Tetrahydrofuran (THF) (8 mL). NH₃ gas wasbubbled in (the mixture changed color yellow to white) and the mixturewas stirred at rt for 15 minutes and then partitioned between brine (15mL) and EtOAc (20 mL). The aqueous phase was extracted with EtOAc (2×20mL) and CH₂Cl₂ (10 mL). The organic phases were combined, dried overNa₂SO₄ and concentrated. The crude product was suspended inN,N-dimethylformamide dimethyl acetal (10 mL, 74.7 mmol) and stirred at105° C. for 2 h. The excess solvent was evaporated and the residue wassuspended in Acetic Acid (10 mL). After the addition of hydrazinemonohydrate (0.194 mL, 3.97 mmol), the mixture was stirred at 100° C.for 1.5 h. The solvent was evaporated and the residue was dissolved inwarm DMSO (8 mL) and purified by reverse phase-HPLC (20-90% AcCN inwater plus 0.1% TFA) to give the desired product (96 mg, 0.197 mmol,19.90% yield) as a white powder. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm13.08 (br. s., 1H), 8.11 (s, 1H), 7.98 (d, J=2.27 Hz, 1H), 7.36 (d,J=8.08 Hz, 1H), 7.01 (t, J=7.96 Hz, 1H), 6.51 (d, J=2.02 Hz, 1H), 6.32(d, J=7.83 Hz, 1H), 5.51 (s, 2H), 3.73-3.97 (m, 4H), 3.12-3.30 (m, 4H),2.50 (s, 3H). MS (ES+) m/e 476.9 [M+H]⁺.

Preparation of2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-4-(1H-tetrazol-5-yl)-1H-benzimidazole

In a two 5 mL MW vial was added in2-methyl-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carbonitrile,prepared as described in Example 24 (80 mg, 0.209 mmol), sodium azide(109 mg, 1.673 mmol) and ammonium chloride (90 mg, 1.673 mmol) andN,N-Dimethylformamide (DMF) (2 mL). The reaction mixture was subjectedto MW irradiation for 15 min at 180° C., then 80 min at 185° C. LC-MSanalysis only showed 30% conversion; however, heating for longer timecaused the decomposition of the product. The two reaction mixtures werecombined. The combined mixture was added in water (10 mL) and extractedwith DCM (30 mL×4). The combined organic phases were washed withsaturated NH₄Cl solution, dried, and concentrated. The reaction wassubjected to purification on a silica column (20˜60% EtOAc/Hexane) andthen (1˜5% MeOH/DCM) to give the product (24 mg, 13%). ¹H NMR (400 MHz,DMSO-d₆) δ ppm 2.53 (s, 3H) 3.08-3.15 (m, 4H) 3.68-3.75 (m, 4H) 6.10 (s,2H) 6.40 (d, J=7.33 Hz, 1H) 7.26-7.39 (m, 2H) 7.56-7.74 (m, 3H) 7.88 (d,J=8.34 Hz, 1H) 8.03 (d, J=8.08 Hz, 1H) 8.26 (d, 1H). MS (ES+) m/e 426.0[M+H]⁺.

Preparation of1-[(3-chloro-2-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added1-(bromomethyl)-3-chloro-2-methylbenzene (0.239 g, 1.090 mmol) andpotassium carbonate (0.301 g, 2.179 mmol). The resulting reactionmixture was stirred for 3 h at 80° C. The solution was cooled to roomtemperature and poured into water and extracted with EtOAc. The combinedorganic phase was washed with brine and concentrated. The residue waspurified on a Biotage Isolera purification system using a Biotage 10 gSNAP silica gel cartridge and eluted with a gradient of DCM to 5%MeOH/DCM over 10 column volumes. The expected compound was collected andevaporated to yield a tan solid. The tan solid was dissolved intetrahydrofuran (THF) (10.00 mL) followed by the addition of 1M lithiumhydroxide solution (10 mL, 10 mmol). The reaction was stirred at 50° C.for 2 h. The reaction was cooled to room temperature and the organicsolvent was removed in-vacuo. The solution was diluted with water (20mL) and acidified with 1 N HCl. The mixture was then filtered and theyellow solid was purified by reversed phase with a gradient ofacetonitrile (0.1% TFA) and water (0.1% TFA v/v)(25-55%) over 10minutes. The appropriate fractions were collected and evaporated toyield the desired product (104.4 mg, 0.253 mmol, 34.9% yield). ¹H NMR(400 MHz, CHLOROFORM-d) δ ppm 2.48 (s, 3H) 2.85 (s, 3H) 3.13 (d, J=4.04Hz, 4H) 3.78-3.90 (m, 4H) 5.58 (s, 2H) 6.37 (d, J=7.83 Hz, 1H) 6.89 (d,J=1.52 Hz, 1H) 7.04 (t, J=7.96 Hz, 1H) 7.37 (d, J=8.08 Hz, 1H) 7.52 (d,1H). MS (ES+) m/e 399.8 [M+H]⁺.

Preparation of2-methyl-1-[(2-methylphenyl)methyl]-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added 2-methylbenzyl bromide(0.145 mL, 1.090 mmol) and potassium carbonate (0.301 g, 2.179 mmol).The resulting reaction mixture was stirred for 3 h at 80° C. Thesolution was cooled to room temperature and poured into water and wasextracted with EtOAc. The combined organic phase was washed with Brineand concentrated. The residue was purified on a Biotage Isolerapurification system using a Biotage 10 g SNAP silica gel cartridge andeluted with a gradient of DCM to 5% MeOH/DCM over 10 column volumes. Theexpected compound was collected and evaporated to yield a tan solid. Thetan solid was dissolved in tetrahydrofuran (THF) (10.00 mL) followed bythe addition of 1M lithium hydroxide solution (10 mL, 10 mmol). Thereaction was stirred at 50° C. for 2 h. The reaction was cooled to roomtemperature and the organic solvent was removed in-vacuo. The solutionwas diluted with water (20 mL) and acidified with 1 N HCl. The mixturewas then filtered and the grey solid was purified by reversed phase HPLCwith a gradient of acetonitrile (0.1% TFA) and water (0.1% TFAv/v)(10-40%) over 10 minutes. The appropriate fractions were collectedand evaporated to yield the desired product (152.6 mg, 0.418 mmol, 57.5%yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 2.45 (s, 3H) 2.87 (s, 3H)3.00 (br. s., 4H) 3.72-3.81 (m, 4H) 5.53 (s, 2H) 6.60 (d, J=7.58 Hz, 1H)6.78 (d, J=1.26 Hz, 1H) 7.14 (t, 1H) 7.23-7.33 (m, 3H). MS (ES+) m/e365.8 [M+H]⁺.

Preparation of ethyl2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate

To a mixture of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid, prepared as described in Example 31 (0.25 g, 0.577 mmol) inDichloromethane (DCM) (20 mL) was added in oxalyl chloride (0.202 mL,2.307 mmol) then followed by ten drops of DMF. The reaction was stirredat rt for 10 minutes and concentrated to give the acid chloride. To thecrude acid chloride, was added Ethanol (20.00 mL). The reaction mixturewas stirred at rt for 10 minutes. The reaction was concentrated. Thecrude product was purified on a silica column (0-10% MeOH/DCM). Thefractions were concentrated and DCM (50 mL) was added in. The organicphase was washed with saturated NaHCO₃ solution (20 mL), Brine (20 mL),dried (MgSO₄) and concentrated to give the product as a white solid(0.18 g, 64%). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.38 (t, J=7.07 Hz, 3H)2.56 (s, 3H) 2.59 (s, 3H) 3.10-3.18 (m, 4H) 3.70-3.78 (m, 4H) 4.45 (q,J=7.07 Hz, 2H) 5.71 (s, 2H) 6.49 (d, J=7.58 Hz, 1H) 7.26 (t, J=7.96 Hz,1H) 7.43 (d, J=2.02 Hz, 1H) 7.55 (d, J=1.52 Hz, 1H) 7.64 (d, 1H). MS(ES+) m/e 462.2[M+H]⁺.

Preparation of 4-bromo-2-methyl-6-(4-morpholinyl)-1H-benzimidazole a)6-bromo-2-methyl-4-nitro-1-(phenylmethyl)-1H-benzimidazole

A mixture of 6-bromo-2-methyl-4-nitro-1H-benzo[d]imidazole, prepared asdescribed in Example 1 (22 g), (bromomethyl)benzene (15 g) and K₂CO₃ (35g) in DMF (250 mL) was stirred at 60° C. for 2 h. The reaction mixturewas cooled to room temperature and filtered. The filtrate was thenpoured into water. It was then filtered to afford a solid and the solidwas washed with water and then dried in-vacuo to afford the desiredproduct (28 g, 93%). ¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.60 (s, 3H), 5.62(s, 2H), 7.12-7.15 (m, 2H), 7.29-7.39 (m, 3H), 8.12 (d, 1H, J=1.8 Hz),8.32 (d, 1H, J=1.8 Hz); LC-MS: m/e=346 [M+1]⁺.

b) 2-methyl-6-(4-morpholinyl)-4-nitro-1-(phenylmethyl)-1H-benzimidazole

A mixture of 6-bromo-2-methyl-4-nitro-1-(phenylmethyl)-1H-benzimidazole(28 g), morpholine (21 g), Pd(dba)₂ (4.6 g), Cs₂CO₃ (52.8 g) and X-Phos(3.9 g) in dioxane (250 mL) was degassed with nitrogen and then stirredat 82° C. for 4 h. The mixture was cooled to room temperature and thesolvent was removed in-vacuo. The residue was then purified by silicagel chromatography eluted with EtOAc:petroleum ether=1:1 to afford thedesired product as a yellow solid (17 g, 60%). ¹H NMR (300 MHz,DMSO-d₆): δ ppm 2.51 (s, 3H), 3.17 (t, 4H, J=4.8 Hz). 3.76 (t, 4H, J=4.8Hz), 5.55 (s, 2H), 7.10-7.13 (m, 2H), 7.28-7.37 (m, 3H), 7.57-7.61 (m,2H); LC-MS: m/e=353 [M+1]⁺.

b) 2-methyl-6-(4-morpholinyl)-1-(phenylmethyl)-1H-benzimidazol-4-amine

To a solution of2-methyl-6-(4-morpholinyl)-4-nitro-1-(phenylmethyl)-1H-benzimidazole (17g) in EtOH (300 mL) was added watery Pd/C (8.7 g) and the mixture wasstirred at 60° C. for 50 h under H₂ atmosphere (4 atm). The mixture wascooled to room temperature and filtered; the filtrate was concentratedin-vacuo. The residue was purified by silica gel chromatography elutedwith EtOAc to afford the desired product as a white solid (7.4 g, 66%).¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.37 (s, 3H), 2.96 (t, 4H, J=4.8 Hz),3.72 (t, 4H, J=4.8 Hz), 4.94 (br s, 2H), 6.04-6.11 (m, 2H), 11.57 (br s,1H); LC-MS: m/e=233 [M+1]⁺

c) 4-bromo-2-methyl-6-(4-morpholinyl)-1H-benzimidazole

To a solution of2-methyl-6-(4-morpholinyl)-1-(phenylmethyl)-1H-benzimidazol-4-amine (2.3g, 10 mmol) in aqueous HBr (50 mL) was added a solution of NaNO₂ (720mg, 10.5 mmol) in water (10 mL) dropwise at 0-5° C. After addition themixture was stirred at 0° C. for minutes, another solution of NaBr (3.1g, 30 mmol) in aqueous HBr (50 mL) was added dropwise at 60° C. Theresulting mixture was then heated to 80° C. for 30 minutes and thencooled to room temperature. It was neutralized with aqueous 2N NaOH andextracted with EtOAc (100 mL×3). The combined organic layers wereconcentrated in-vacuum and the residue was purified by silica gelchromatography eluted with petroleum ether:EtOAc=1:1 to give the desiredproduct (1.7 g, 58%) as a white solid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm2.44 (s, 3H), 3.07 (t, 4H, J=4.8 Hz), 3.74 (t, 4H, J=4.8 Hz), 6.85 (d,1H, J=1.8 Hz), 7.04 (d, 1H, J=1.8 Hz), 12.20 (br s, 1H); LC-MS: m/e=296[M+1]⁺

Preparation of4-bromo-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole

A suspension of 4-bromo-2-methyl-6-(4-morpholinyl)-1H-benzimidazole,prepared as described in Example 62 (500 mg, 1.688 mmol) and potassiumcarbonate (700 mg, 5.06 mmol) in N,N-Dimethylformamide (DMF) (6 mL) wasstirred at rt for 15 min.1-(bromomethyl)-2-methyl-3-(trifluoromethyl)benzene (641 mg, 2.53 mmol)was added in and the resulting reaction mixture was stirred for 3 h at80° C. It was then cooled to room temperature and poured into ice/water.The precipitate was collected by filtration, washed with water, then fewmLs of hexanes and air dried. The crude material was purified on asilica gel column (ISCO, 0-80% EtOAc in hexanes) to the desired product(565 mg, 1.182 mmol, 70.0% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm7.59 (d, J=7.83 Hz, 1H), 7.16 (d, J=2.02 Hz, 1H), 7.10-7.15 (m, 1H),6.46-6.52 (m, 2H), 5.25 (s, 2H), 3.75-3.89 (m, 4H), 3.05-3.14 (m, 4H),2.55 (s, 3H), 2.51 (s, 3H). MS (ES+) m/e 468.9 [M+H]⁺.

Preparation of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1,3-oxazol-2-yl)-1H-benzimidazole

A mixture of4-bromo-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole(150 mg, 0.320 mmol), prepared as described in Example 63,2-(tributylstannanyl)-1,3-oxazole (195 mg, 0.545 mmol) and Pd(Ph₃P)₂Cl₂(11.27 mg, 0.016 mmol) in Tetrahydrofuran (THF) (5 mL) was stirred atreflux temperature for 19 h. Conversion to the desired product isobserved by LC/MS analysis, but the majority of the mixture is still SM.The reaction mixture was transferred in a microwavable vial andirradiated in a microwave reactor at 120° C. for 90 min. The reactionmixture was diluted with EtOAc and CHCl₃, washed with aq sat sol NH₄Cl,brine, dried over Na₂SO₄ and the solvent was evaporated under reducedpressure. The residue was purified on a silica gel column (ISCO, 0-70%EtOAc in Hexanes—no product peak observed—then 0-10% MeOH in CH₂Cl₂) togive desired product (94.8 mg, 0.204 mmol, 63.5% yield) as a yellowpowder. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.87 (s, 1H), 7.71 (d,J=2.27 Hz, 1H), 7.58 (d, J=7.83 Hz, 1H), 7.37 (s, 1H), 7.11 (t, J=7.83Hz, 1H), 6.66 (d, J=2.27 Hz, 1H), 6.47 (d, J=7.83 Hz, 1H), 5.31 (s, 2H),3.81-3.92 (m, 4H), 3.11-3.24 (m, 4H), 2.58 (s, 3H), 2.56 (s, 3H). MS(ES+) m/e 457.1 [M+H]⁺

Example 65

Preparation of methyl2-chloro-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate a) methyl6-(4-morpholinyl)-2-oxo-2,3-dihydro-1H-benzimidazole-4-carboxylate

To a solution of methyl 2,3-diamino-5-(4-morpholinyl)benzoate, preparedas described in Example 45, step a (11.0 g, 4.0 mmol) in DMF (50 mL) wasadded urea (720 mg, 12 mmol) and the mixture was heated to 170° C. for 4h. When analysis by TLC showed no starting material remaining, themixture was cooled to room temperature then diluted with DCM (200 mL),washed with water (50 mL×2) and dried over anhydrous Na₂SO₄, filteredand concentrated in-vacuo. The residue was then purified bychromatography on silica (eluted with EtOAc) to afford the desiredproduct as a dark-yellow solid (690 mg, 62%). ¹H NMR (300 MHz, DMSO-d₆):δ ppm 3.02 (t, 4H, J=4.8 Hz), 3.74 (t, 4H, J=4.8 Hz), 3.86 (s, 3H), 6.82(d, 1H, J=2.1 Hz), 6.99 (d, 1H, J=2.1 Hz), 10.48 (s, 1H), 10.82 (s, 1H).LC-MS: m/e=278 [M+1]⁺.

b) methyl 2-chloro-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate

To a solution of combined batches of methyl6-(4-morpholinyl)-2-oxo-2,3-dihydro-1H-benzimidazole-4-carboxylate (6.8g, 24.5 mmol) in POCl₃ (25 mL) was added N,N-Dimethylaniline (8.8 g,73.5 mmol) and the mixture was heated to 103° C. for 12 h. When TLCanalysis showed no starting material remaining, the mixture was cooledto room temperature, purified by chromatography (eluted with petroleumether/EtOAc=1/1) on silica to afford the desired product as a off-whitesolid (1.6 g, 23%). ¹H NMR (300 MHz, DMSO-d₆): δ ppm 3.11 (t, 4H, J=4.5Hz), 3.77 (t, 4H, J=4.5 Hz), 3.93 (s, 3H), 7.43 (d, 1H, J=1.8 Hz), 7.50(d, 1H, J=1.8 Hz), 12.97 (s, 1H). LC-MS: m/e=296 [M+1]⁺.

Preparation of methyl2-chloro-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate

To the mixture of methyl2-chloro-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 65 (0.5 g, 1.691 mmol) in N,N-Dimethylformamide(DMF) (10 ml) was added in potassium carbonate (0.467 g, 3.38 mmol) and1-(bromomethyl)-2-methyl-3-(trifluoromethyl)benzene (0.428 g, 1.691mmol). The reaction mixture was stirred at 80° C. for 1 h. The reactionwas cooled down. Water (100 mL) was added in. The solid precipitated.Filtration gave the solid which was purified on a silica column (20˜60%EtOAc/Hexane) to give the product as white solid (0.66 g, 83%). ¹H NMR(400 MHz, DMSO-d₆) δ ppm 2.54 (s, 3H) 3.07-3.16 (m, 4H) 3.68-3.78 (m,4H) 3.91 (s, 3H) 5.63 (s, 2H) 6.41 (d, J=7.83 Hz, 1H) 7.28 (t, J=7.83Hz, 1H) 7.39 (d, J=2.27 Hz, 1H) 7.50 (d, J=2.53 Hz, 1H) 7.63 (d, 1H). MS(ES+) m/e 468.0 [M+H]⁺.

Preparation of2-chloro-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl2-chloro-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylate,prepared as described in Example 66 (0.6 g, 1.282 mmol) inTetrahydrofuran (THF) (10 mL) was added in 2 N lithium hydroxide (6.41mL, 12.82 mmol). The reaction mixture was stirred at 50 C for 70 min.The reaction was cooled down. The organic solvent was removed in-vacuo.The aqueous mixture was acidified using 1 N HCl. The solid precipitated.Filtration and washing with water gave the product as white solid (0.55g, 90%). ¹H NMR (400 MHz, DMSO-d₆) d ppm 2.54 (s, 3H) 3.05-3.16 (m, 4H)3.67-3.78 (m, 4H) 5.63 (s, 2H) 6.43 (d, J=7.83 Hz, 1H) 7.28 (t, J=7.83Hz, 1H) 7.36 (d, J=2.27 Hz, 1H) 7.49 (d, J=2.53 Hz, 1H) 7.63 (d, J=8.08Hz, 1H) 12.90 (s, 1H). MS (ES+) m/e 453.9 [M+H]⁺.

Preparation of methyl2-(difluoromethyl)-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate a)methyl 2,3-diamino-5-morpholinobenzoate

To a mixture of intermediate methyl3-amino-5-(4-morpholinyl)-2-nitrobenzoate, prepared as described inExample 26, step c (98 g, 0.35 mol) in MeOH (2.2 L) was added Pd/C (9.8g, 10%) and the resulting mixture was then stirred at room temperatureunder H₂ (4 atm) atmosphere. After stirring for 16 h, it was filteredand concentrated in vacuum to give the crude product (84.4 g, 96%) as adark solid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.85 (t, 4H, J=4.8 Hz),3.70 (t, 4H, J=4.8 Hz), 3.76 (s, 3H), 4.77 (br. S, 2H), 5.86 (br. S,2H), 6.54 (d, 1H, J=2.7 Hz), 6.61 (d, 1H, J=2.7 Hz); LC-MS: m/e=252[M+1]⁺

b) methyl2-(difluoromethyl)-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate

A mixture of methyl 2,3-diamino-5-morpholinobenzoate (40.16 g, 160 mmol)and 2,2-difluoroacetic acid (46.08 g, 480 mmol) in toluene (500 mL) wasstirred at reflux temperature for 15 h. Then the mixture was cooled toroom temperature and the solvent was removed in vacuum. The residue waspurified by silica gel chromatography eluted with petroleumether:EtOAc=2:1 to afford the desired product. Then, it was dissolvedwith EtOAc (2 L) and washed with aqueous NaHCO₃ (1 L) and brine (1 L).The organic layer was dried over anhydrous Na₂SO₄ and concentrated togive the desired product (40.9 g, 82%) as a yellow solid. ¹H NMR showedthis compound is in a form of tautomeric mixture (major tautomer/minortautomer=5/1) ¹H NMR of the major tautomer (300 MHz, DMSO-d₆): δ ppm3.14 (t, 4H, J=4.8 Hz), 3.78 (t, 4H, J=4.8 Hz), 3.96 (s, 3H), 7.21 (t,1H, J=52.8 Hz), 7.55 (d, 1H, J=2.4 Hz), 7.65 (d, 1H, J=2.4 Hz), 12.92(s, 1H); LC-MS: m/e=312 [M+1]⁺

Preparation of2-(difluoromethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid a) methyl2-(difluoromethyl)-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazole-4-carboxylate

A mixture of methyl2-(difluoromethyl)-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate,prepared as described in Example 68 (500 mg, 1.6 mmol), K₂CO₃ (442 mg,3.2 mmol) and 1-(bromomethyl)naphthalene (426 mg, 1.9 mmol) in DMF (15mL) was stirred at 70° C. for 18 h. The reaction mixture was cooled toroom temperature and filtered. The liquid was poured into water (100 mL)and filtered, the filter cake was collected and purified by silica gelchromatography eluted with petroleum ether:EtOAc=1:1 to give the desiredproduct (710 mg, 98%) as a yellow solid. ¹H NMR (300 MHz, DMSO-d₆): δppm 3.08 (t, 4H, J=4.5 Hz), 3.68 (t, 4H, J=4.5 Hz), 3.94 (s 3H), 6.21(s, 2H), 6.27 (d, 1H, J=6.9 Hz), 7.21-7.38 (m, 3H), 7.56-7.69 (m, 3H),7.84 (d, 1H, J=8.4 Hz), 8.00 (d, 1H, J=8.4 Hz), 8.24 (d, 1H, J=8.7 Hz);LC-MS: m/e=452 [M+1]⁺

b)2-(difluoromethyl)-6-(4-morpholinyl)-1-(1-naphthalenylmethyl)-1H-benzimidazole-4-carboxylicacid

A mixture methyl2-(difluoromethyl)-6-morpholino-1-(naphthalen-1-ylmethyl)-1H-benzo[d]imidazole-4-carboxylate(700 mg, 1.55 mmol) and 2N LiOH (5 mL) in THF (10 mL) was stirred at 45°C. for 4 h. It was filtered, the filter cake was dissolved in water (10mL) and formic acid was added to adjust pH to 3-4. Then a filtration wasperformed and the filter cake was collected, dried under vacuum to givethe product (450 mg, 66%) as a white solid. ¹H NMR (300 MHz, DMSO-d₆): δppm 3.07 (s, 4H), 3.68 (s, 4H, s), 6.20 (s, 2H), 6.30 (d, 1H, J=7.2 Hz),7.20-7.72 (m, 6H), 7.85 (d, 1H, J=8.1 Hz), 8.01 (d, 1H, J=7.8 Hz), 8.24(d, 1H, J=7.5 Hz); LC-MS: m/e=438 [M+1]⁺

Example 70

Preparation of2-(difluoromethyl)-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid a) methyl2-(difluoromethyl)-1-(2-methyl-3-(trifluoromethyl)benzyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylate

A mixture of methyl2-(difluoromethyl)-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate,prepared as described in Example 68 (500 mg, 1.6 mmol), K₂CO₃ (442 mg,3.2 mmol) and 1-(bromomethyl)-2-methyl-3-(trifluoromethyl)benzene (480mg, 1.9 mmol) in DMF (15 mL) was stirred at 70° C. for 18 h. Thereaction mixture was cooled to room temperature and filtered. Thefiltrate was poured into water (100 mL) and filtered, the filter cakewas collected and purified by silica gel chromatography eluted withpetroleum ether:EtOAc=1:1 to give the desired product (710 mg, 98%) as ayellow solid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm 2.53 (s, 3H), 3.14 (t,4H, J=4.5 Hz), 3.73 (t, 4H, J=4.5 Hz), 3.93 (s, 3H), 5.75 (s, 2H), 6.27(d, 1H, J=7.8 Hz), 7.22 (t, 1H, J=7.8 Hz), 7.30 (d, 1H, J=2.1 Hz), 7.36(t, 1H, J=51.6 Hz), 7.58-7.61 (m, 2H); LC-MS: m/e=484 [M+1]⁺

b)2-(difluoromethyl)-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl2-(difluoromethyl)-1-(2-methyl-3-(trifluoromethyl)benzyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylate(700 mg, 1.45 mmol) and 2 N LiOH (5 mL) in THF (10 mL) was stirred at45° C. for 4 h. It was filtered, the filter cake was dissolved in water(10 mL) and formic acid was added to adjust pH=3-4. Then a filtrationwas performed and the filter cake was dried under vacuum to give thedesired product (400 mg, 59%) as a white solid. ¹H NMR (300 MHz,DMSO-d₆): δ ppm 2.53 (s, 3H), 3.13 (s, 4H), 3.73 (s, 4H), 5.75 (s, 2H),6.29 (d, 1H, J=7.5 Hz), 7.19-7.61 (m, 5H), 12.97 (br s, 1H); LC-MS:m/e=470 [M+1]⁺

Preparation of1-[(2,3-dichlorophenyl)methyl]-2-(difluoromethyl)-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid a) methyl1-(2,3-dichlorobenzyl)-2-(difluoromethyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylate

A mixture of methyl2-(difluoromethyl)-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate,prepared as described in Example 68 (1000 mg, 3.2 mmol), K₂CO₃ (884 mg,6.4 mmol) and 1-(bromomethyl)-2,3-dichlorobenzene (926 mg, 3.8 mmol) inDMF (30 mL) was stirred at 70° C. for 18 h. The reaction mixture wascooled to room temperature and filtered. The filtrate was poured intowater (100 mL) and filtered, the filter cake was collected and purifiedby silica gel chromatography eluted with petroleum ether:EtOAc=1:1 togive the desired product (1.4 g, 93%) as a yellow solid. ¹H NMR (300MHz, DMSO-d₆): δ ppm 3.16 (s, 4H), 3.74 (s, 4H), 3.92 (s, 3H), 5.77 (s,2H), 6.21 (d, 1H, J=7.5 Hz), 7.20-7.58 (m, 5H); LC-MS: m/e=470 [M+1]⁺

b)1-[(2,3-dichlorophenyl)methyl]-2-(difluoromethyl)-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

A mixture of methyl1-(2,3-dichlorobenzyl)-2-(difluoromethyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylate(1350 mg, 2.88 mmol) and 2 N LiOH (10 mL) in THF (20 mL) was stirred at45° C. for 4 h. It was filtered, the filter cake was dissolved in water(10 mL) and formic acid was added to adjust pH=3-4. Then a filtrationwas performed and the filter cake was dried under vacuum to give thedesired product (600 mg, 46%) as a white solid. ¹H NMR (300 MHz,DMSO-d₆): δ ppm 3.15 (s, 4H), 3.73 (s, 4H), 5.76 (s, 2H), 6.22 (d, 1H,J=7.5 Hz), 7.20-7.60 (m, 5H), 12.98 (br s, 1H); LC-MS: m/e=456 [M+1]⁺

Preparation of1-[(3-chloro-2-methylphenyl)methyl]-2-(difluoromethyl)-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid a) methyl1-(2,3-dichlorobenzyl)-2-(difluoromethyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylate

A mixture of methyl2-(difluoromethyl)-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate,prepared as described in Example 68 (1.18 g, 3.8 mmol), K₂CO₃ (2.48 g,7.6 mmol) and 1-(bromomethyl)-3-chloro-2-methylbenzene (1 g, 4.6 mmol)in DMF (40 mL) was stirred at 70° C. for 18 h. The reaction mixture wascooled to room temperature and filtered. The filtrate was poured intowater (100 mL) and filtered, the filter cake was collected and purifiedby silica gel chromatography eluted with petroleum ether:EtOAc=1:1 togive the desired product (1.18 g, 69%) as a yellow solid. ¹H NMR (300MHz, DMSO-d₆): δ ppm 2.46 (s, 3H), 3.13 (t, 4H, J=4.8 Hz), 3.73 (t, 4H,J=4.8 Hz), 3.92 (s, 3H), 5.70 (s, 2H), 5.97 (d, 1H, J=7.5 Hz), 7.04 (t,1H, J=7.5 Hz), 7.26 (d, 1H, J=1.8 Hz), 7.34 (t, 1H, J=7.5 Hz), 7.35 (t,1H, J=51.6 Hz), 7.58 (d, 1H, J=1.8 Hz); LC-MS: m/e=450 [M+1]⁺

b)1-[(3-chloro-2-methylphenyl)methyl]-2-(difluoromethyl)-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

A solution of methyl1-(2,3-dichlorobenzyl)-2-(difluoromethyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylate(1045 mg, 2.3 mmol) in THF (40 mL) was added into 2 N LiOH (20 mL) andthe mixture was stirred at 45° C. for 4 h. It was filtered, the filtercake was added to water (100 mL) and formic acid was added to adjustpH=3. Then a filtration was performed, the filter cake was collected andwashed with water (200 mL), dried under vacuum to give the desiredproduct (800 mg, 80%) as a yellow solid. ¹H NMR (300 MHz, DMSO-d₆): δppm 2.46 (s, 3H), 3.13 (t, 4H, J=4.8 Hz), 3.73 (t, 4H, J=4.8 Hz), 3.92(s, 3H), 5.70 (s, 2H), 6.00 (d, 1H, J=7.5 Hz), 7.04 (t, 1H, J=7.5 Hz),7.23 (d, 1H, J=1.8 Hz), 7.34 (t, 1H, J=7.5 Hz), 7.36 (t, 1H, J=51.9 Hz),7.57 (d, 1H, J=1.8 Hz), 12.96 (br s, 1H); LC-MS: m/e=436 [M+1]⁺

Example 73

Preparation of1-(1-benzothien-7-ylmethyl)-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added7-(bromomethyl)-1-benzothiophene (0.247 g, 1.090 mmol) and potassiumcarbonate (0.301 g, 2.179 mmol). The resulting reaction mixture wasstirred for 3 h at 80° C. An additional amount of7-(bromomethyl)-1-benzothiophene (0.247 g, 1.090 mmol) was added andmixture was stirred for 3 h at 80° C. The solution was cooled to roomtemperature and poured into water and extracted with EtOAc. The combinedorganic phase was washed with brine and concentrated. The residue waspurified on Biotage Isolera purification system using a Biotage 10 gSNAP silica gel cartridge and eluted with a gradient of DCM to 5%MeOH/DCM over 10 column volumes. The expected compound was collected andevaporated to yield a tan solid. The tan solid was dissolved intetrahydrofuran (THF) (10.00 mL) followed by the addition of 1M lithiumhydroxide solution (10 mL, 10 mmol). The reaction was found to beincomplete so the solution was neutralized with 1M HCl and evaporated.The residue was dissolved in 5 mL of methanol and treated with 1N NaOHfor 2 h at 50° C., which resulted in a complete reaction. The reactionwas cooled to room temperature and the organic solvent was removed invacuo. The solution was diluted with water (20 mL) and acidified with 1N HCl. The mixture was then filtered and the yellow solid was isolated.The aqueous layer was found to contain a significant amount of productand was evaporated. Both solid and residue were purified by reversedphase chromatography with a gradient of acetonitrile (0.1% TFA) andwater (0.1% TFA v/v) (10-45%) over 10 minutes. The appropriate fractionswere collected and evaporated to the desired product (27.9 mg, 0.068mmol, 9.42% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.76 (s, 3H)3.14-3.21 (m, 4H) 3.71-3.76 (m, 4H) 6.01 (s, 2H) 7.06 (d, J=7.07 Hz, 1H)7.36-7.44 (m, 1H) 7.56-7.62 (m, 2H) 7.71 (d, J=2.27 Hz, 1H) 7.83 (d,J=5.31 Hz, 1H) 7.92 (d, 1H). MS (ES+) m/e 408.1 [M+H]⁺.

Preparation of1-[(2,3-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added 2,3-dimethylbenzyl bromide(0.289 g, 1.453 mmol) and potassium carbonate (0.301 g, 2.179 mmol). Theresulting reaction mixture was stirred for 3 h at 80° C. The solutionwas cooled to room temperature and poured into water and was extractedwith EtOAc. The combined organic phase was washed with Brine andconcentrated. The residue was purified on Biotage Isolera purificationsystem using a Biotage 10 g SNAP silica gel cartridge and eluted with agradient of DCM to 5% MeOH/DCM over 10 column volumes. The expectedcompound was collected and evaporated to yield a tan solid. The tansolid was dissolved in tetrahydrofuran (THF) (10.00 mL) followed by theaddition of 1M lithium hydroxide solution (10 mL, 10 mmol). The reactionwas stirred at 50° C. for 2 h. The reaction was cooled to roomtemperature and the organic solvent was removed in vacuo. The solutionwas diluted with water (20 mL) and acidified with 1 N HCl. The mixturewas then filtered and the grey solid was purified by reversed phase HPLCwith a gradient of acetonitrile (0.1% TFA) and water (0.1% TFA v/v)(20-50%) over 10 minutes. The appropriate fractions were collected andevaporated to yield the desired product (55.2 mg, 0.145 mmol, 20.02%yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 2.31 (s, 3H) 2.36 (s, 3H)2.85 (s, 3H) 3.13-3.24 (m, 4H) 3.80-3.96 (m, 4H) 5.49-5.61 (m, 2H)6.29-6.38 (m, 1H) 6.94-6.99 (m, 1H) 7.00-7.07 (m, 1H) 7.14-7.23 (m, 1H)7.72 (m, 1H). MS (ES+) m/e 379.8 [M+H]⁺.

Preparation of1-[(3-fluoro-2-methylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added1-(bromomethyl)-3-fluoro-2-methylbenzene (0.295 g, 1.453 mmol) andpotassium carbonate (0.301 g, 2.179 mmol). The resulting reactionmixture was stirred for 3 h at 80° C. The solution was cooled to roomtemperature and poured into water and was extracted with EtOAc. Thecombined organic phase was washed with Brine and concentrated. Theresidue was purified on Biotage Isolera purification system using aBiotage 10 g SNAP silica gel cartridge and eluted with a gradient of DCMto 5% MeOH/DCM over 10 column volumes. The expected compound wascollected and evaporated to yield a tan solid. The tan solid wasdissolved in tetrahydrofuran (THF) (10.00 mL) followed by the additionof 1M lithium hydroxide solution (10 mL, 10 mmol). The reaction wasstirred at 50° C. for 2 h. The reaction was cooled to room temperatureand the organic solvent was removed in vacuo. The solution was dilutedwith water (20 mL) and acidified with 1 N HCl. The mixture was thenfiltered and the grey solid was purified by reversed phase HPLC with agradient of acetonitrile (0.1% TFA) and water (0.1% TFA v/v) (10-40%)over 10 minutes. The appropriate fractions were collected and evaporatedto yield the desired product (62.7 mg, 0.164 mmol, 22.51% yield). ¹H NMR(400 MHz, CHLOROFORM-d) δ ppm 2.36 (s, 3H) 2.87 (s, 3H) 3.07-3.20 (m,4H) 3.81-3.92 (m, 4H) 5.55 (s, 2H) 6.29-6.38 (m, 1H) 6.85-6.90 (m, 1H)7.03-7.17 (m, 2H) 7.55 (m, 1H). MS (ES+) m/e 383.8 [M+H]⁺.

Preparation of2,4-dimethyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole

A mixture of4-bromo-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole,prepared as described in Example 62 (200 mg, 0.427 mmol),trimethylboroxine (0.239 mL, 1.708 mmol), Pd(Ph₃P)₄ (49.4 mg, 0.043mmol) and potassium carbonate (118 mg, 0.854 mmol) in 1,4-Dioxane (2.5mL)/Water (0.25 mL) was irradiated at 120° C. in a microwave synthesizerfor 40 min, then cooled and poured into water. The mixture was extractedwith ethyl acetate. The extracts were washed with brine, dried (Na₂SO₄)and evaporated under reduced pressure. The residue was purified byRP-HPLC (25-45% AcCN in water plus 0.1% TFA) to give the desiredcompound (77 mg, 0.181 mmol, 42.5% yield) as a white solid (contains3-5% of the 4-H compound). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.58 (d,J=8.08 Hz, 1H), 7.12 (t, J=7.83 Hz, 1H), 6.80 (d, J=1.01 Hz, 1H), 6.51(d, J=7.83 Hz, 1H), 6.40 (d, J=1.77 Hz, 1H), 5.26 (s, 2H), 3.78-3.91 (m,4H), 3.01-3.15 (m, 4H), 2.67 (s, 3H), 2.56 (s, 3H), 2.50 (s, 3H). MS(ES+) m/e 404.1 [M+H]⁺. (NOTE: The reaction was repeated usingPdCl₂(dppf) as catalyst. Less (to negligible) reduction was observed).

Preparation of1-[1-(3-chloro-2-methylphenyl)ethyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added1-(1-bromoethyl)-3-chloro-2-methylbenzene (0.339 g, 1.453 mmol) andpotassium carbonate (0.301 g, 2.179 mmol). The resulting reactionmixture was stirred for 3 h at 80° C. The solution was cooled to roomtemperature and poured into water and was extracted with EtOAc. Thecombined organic phase was washed with Brine and concentrated. Theresidue was purified on Biotage Isolera purification system using aBiotage 10 g SNAP silica gel cartridge and eluted with a gradient of DCMto 5% MeOH/DCM over 10 column volumes. The expected compound wascollected and evaporated to yield a tan solid. The tan solid wasdissolved in tetrahydrofuran (THF) (10.00 mL) followed by the additionof 1M lithium hydroxide solution (10 mL, 10 mmol). The reaction wasstirred at 50° C. for 2 h. The reaction was cooled to room temperatureand the organic solvent was removed in vacuo. The solution was dilutedwith water (20 mL) and acidified with 1 N HCl. The mixture was thenfiltered and the grey solid was purified by reversed phase HPLC with agradient of acetonitrile (0.1% TFA) and water (0.1% TFA v/v) (20-50%)over 10 minutes. The appropriate fractions were collected and evaporatedto yield the desired product (36.1 mg, 0.087 mmol, 12.01% yield). ¹H NMR(400 MHz, CHLOROFORM-d) δ ppm 2.04-2.10 (m, 6H) 2.88 (s, 3H) 2.98-3.17(m, 4H) 3.87 (s, 4H) 5.95-6.06 (m, 1H) 6.70-6.77 (m, 1H) 7.35-7.42 (m,1H) 7.51-7.57 (m, 1H) 7.60-7.67 (m, 1H) 7.71-7.77 (m, 1H). MS (ES+) m/e413.8 [M+H]⁺.

Example 78

Preparation of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-4-(1,3-thiazol-2-yl)-1H-benzimidazole

A mixture of4-bromo-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole,prepared as described in Example 62 (200 mg, 0.427 mmol), 2-thazolylzincbromide (1.708 mL, 0.854 mmol) and Pd(Ph₃P)₄ (49.4 mg, 0.043 mmol) inTetrahydrofuran (THF) (1.5 mL) was irradiated in a microwave reactor at110° C. for 2.5 h. The reaction mixture was diluted with EtOAc andCHCl₃, washed with NH₄Cl aq sat sol, brine, dried over Na₂SO₄ andevaporated under reduced pressure. The residue was purified on silicagel (ISCO, 0-70% EtOAc in Hexanes, then 0-10% MeOH in CH₂Cl₂) to givethe desired product (127 mg, 0.255 mmol, 59.8% yield) as a yellowpowder. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.97 (d, J=3.28 Hz, 1H),7.55 (d, J=3.54 Hz, 1H), 7.45 (d, J=7.83 Hz, 1H), 7.32 (d, J=2.02 Hz,1H), 7.03 (t, J=7.83 Hz, 1H), 6.74 (d, J=1.77 Hz, 1H), 6.35 (d, J=7.83Hz, 1H), 5.32 (s, 2H), 3.60-3.76 (m, 4H), 2.95-3.07 (m, 4H), 2.60 (s,3H), 2.55 (s, 3H). MS (ES+) m/e 473.1 [M+H]⁺.

Preparation of4-(2-furanyl)-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole

A mixture of4-bromo-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole,prepared as described in Example 62 (200 mg, 0.427 mmol),2-furanyl-boronic acid (71.7 mg, 0.641 mmol), PdCl₂(dppf)-CH₂Cl₂ adduct(34.9 mg, 0.043 mmol) and sodium carbonate (91 mg, 0.854 mmol) in1,2-Dimethoxyethane (DME) (2.5 mL) and Water (0.5 mL) was irradiated ina microwave reactor for 1 h at 100° C. The mixture was poured into waterand extracted with EtOAc. The organic phase was washed with brine, driedover Na₂SO₄ and evaporated. The residue was purified by RP-HPLC (Gilson,25-65% Acetonitrile in water plus 0.1% TFA) to give the desired product(48.5 mg, 0.104 mmol, 24.43% yield) as a white powder (separation fromimpurity was difficult. The head of the peak was discarded decreasingthe overall yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.51-7.63 (m,3H), 7.42 (d, J=2.02 Hz, 1H), 7.12 (t, J=7.83 Hz, 1H), 6.60 (dd, J=3.28,1.77 Hz, 1H), 6.51 (d, J=7.83 Hz, 1H), 6.49 (d, J=2.02 Hz, 1H), 5.28 (s,2H), 3.80-3.94 (m, 4H), 3.12-3.22 (m, 4H), 2.56 (s, 3H), 2.53 (s, 3H).MS (ES+) m/e 456.0 [M+H]⁺.

Preparation of2-methyl-4-[(methyloxy)methyl]-1-{[2-methyl-3-[(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole

To the mixture of[2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazol-4-yl]methanol,prepared as described in Example 43 (160 mg, 0.381 mmol) inN,N-Dimethylformamide (DMF) (15 mL), sodium hydride (30.5 mg, 0.763mmol) was added in and followed by the addition of methyl iodide (0.048mL, 0.763 mmol). The reaction was stirred at rt for 3 hours. More sodiumhydride (30.5 mg, 0.763 mmol) and methyl iodide (0.048 mL, 0.763 mmol)was added in. The reaction was stirred at rt for another 2 hours thenWater (70 mL) was added in. The mixture was extracted with EtOAc (100mL). The organic phase was washed with Brine (100 mL), dried (MgSO₄) andconcentrated. The crude was subjected to ISCO purification (0-2%MeOH/DCM) to give the product. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.36 (s,3H) 2.54 (s, 3H) 3.00-3.07 (m, 4H) 3.37 (s, 3H) 3.68-3.75 (m, 4H) 4.76(s, 2H) 5.52 (s, 2H) 6.32 (d, J=7.83 Hz, 1H) 6.88 (m, 2H) 7.25 (s, 1H)7.60 (d, 1H). MS (ES+) m/e 434.4 [M+H]⁺.

Preparation of4-(3-furanyl)-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole

A mixture of4-bromo-2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole,prepared as described in Example 62 (200 mg, 0.427 mmol), 3-furanylboronic acid (47.8 mg, 0.427 mmol), sodium carbonate (91 mg, 0.854 mmol)and PdCl₂(dppf)-CH₂Cl₂ adduct (34.9 mg, 0.043 mmol) in1,2-Dimethoxyethane (DME) (2.5 mL) and Water (0.5 mL) was irradiated ina microwave reactor for 1 h at 100° C. The mixture was poured into waterand extracted with EtOAc. The organic phase was washed with brine, driedover Na₂SO₄ and evaporated. Only partial conversion (ca. 50%) wasobserved by LC/MS analysis. The residue was purified by RP-HPLC (Gilson,25-65% Acetonitrile in water plus 0.1% TFA) to the desired product (28mg, 0.060 mmol, 14.11% yield) as a white powder (separation fromimpurity was difficult. The head of the peak was discarded decreasingthe overall yield, in addition to the partial conversion observed). ¹HNMR (400 MHz, CHLOROFORM-d) δ ppm 8.57 (s, 1H), 7.58 (d, J=7.58 Hz, 1H),7.55 (t, J=1.52 Hz, 1H), 7.09-7.15 (m, 2H), 7.02 (d, J=1.26 Hz, 1H),6.52 (d, J=7.83 Hz, 1H), 6.48 (d, J=2.02 Hz, 1H), 5.29 (s, 2H),3.80-3.97 (m, 4H), 3.07-3.21 (m, 4H), 2.57 (s, 3H), 2.52 (s, 3H). MS(ES+) m/e 456.0 [M+H]⁺

Preparation of2-methyl-1-{[2-methyl-5-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.3 g, 1.090 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added2-methyl-5-(trifluoromethyl)benzyl bromide (0.552 g, 2.179 mmol) andpotassium carbonate (0.452 g, 3.27 mmol). The resulting reaction mixturewas stirred for 3 h at 80° C. The solution was cooled to roomtemperature and poured into water and was extracted with EtOAc. Thecombined organic phase was washed with Brine and concentrated. Theresidue was purified on Biotage Isolera purification system using aBiotage 10 g SNAP silica gel cartridge and eluted with a gradient of DCMto 5% MeOH/DCM over 10 column volumes. The expected compound wascollected and evaporated to yield a tan solid. The tan solid wasdissolved in tetrahydrofuran (THF) (10.00 mL) followed by the additionof 1M lithium hydroxide solution (10 mL, 10 mmol). The reaction wasstirred at 50° C. for 2 h. The reaction was cooled to room temperatureand the organic solvent was removed in vacuo. The solution was dilutedwith water (20 mL) and acidified with 1 N HCl. The mixture was thenfiltered and the grey solid was purified by reversed phase HPLC with agradient of acetonitrile (0.1% TFA) and water (0.1% TFA v/v) (25-55%)over 10 minutes. The appropriate fractions were collected and evaporatedto yield the desired product (120.1 mg, 0.277 mmol, 25.4% yield). ¹H NMR(400 MHz, METHANOL-d₄) δ ppm 2.53 (s, 3H) 2.88 (s, 3H) 3.15-3.23 (m, 4H)3.79-3.88 (m, 4H) 5.84 (s, 2H) 7.06 (s, 1H) 7.27 (d, J=2.27 Hz, 1H)7.51-7.58 (m, 1H) 7.59-7.64 (m, 1H) 7.86 (d, 1H). MS (ES+) m/e 433.8[M+H]⁺.

Preparation of1-[(2,5-dimethylphenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added 2,5-dimethylbenzyl bromide(0.289 g, 1.453 mmol) and potassium carbonate (0.301 g, 2.179 mmol). Theresulting reaction mixture was stirred for 3 h at 80° C. The solutionwas cooled to room temperature and poured into water and was extractedwith EtOAc. The combined organic phase was washed with Brine andconcentrated. The residue was purified on Biotage Isolera purificationsystem using a Biotage 10 g SNAP silica gel cartridge and eluted with agradient of DCM to 5% MeOH/DCM over 10 column volumes. The expectedcompound was collected and evaporated to yield a tan solid. The tansolid was dissolved in tetrahydrofuran (THF) (10.00 mL) followed by theaddition of 1M lithium hydroxide solution (10 mL, 10 mmol). The reactionwas stirred at 50° C. for 2 h. The reaction was cooled to roomtemperature and the organic solvent was removed in vacuo. The solutionwas diluted with water (20 mL) and acidified with 1 N HCl. The mixturewas then filtered and the grey solid was purified by reversed phase HPLCwith a gradient of acetonitrile (0.1% TFA) and water (0.1% TFA v/v)(15-50%) over 10 minutes. The appropriate fractions were collected andevaporated to yield the desired product (96.4 mg, 0.254 mmol, 35.0%yield). ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 2.19 (s, 3H) 2.40 (s, 3H)2.84 (s, 3H) 3.13-3.27 (m, 4H) 3.78-3.86 (m, 4H) 5.70 (s, 2H) 6.54 (s,1H) 7.09 (d, J=7.83 Hz, 1H) 7.20 (d, J=7.58 Hz, 1H) 7.25 (d, J=2.27 Hz,1H) 7.83 (d, 1H). MS (ES+) m/e 379.8 [M+H]⁺.

Example 84

Preparation of1-[1-(3-chlorophenyl)ethyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added1-(1-bromoethyl)-3-chlorobenzene (0.203 mL, 1.453 mmol) and potassiumcarbonate (0.301 g, 2.179 mmol). The resulting reaction mixture wasstirred for 3 h at 80° C. The solution was cooled to room temperatureand poured into water and was extracted with EtOAc. The combined organicphase was washed with Brine and concentrated. The residue was purifiedon Biotage Isolera purification system using a Biotage 10 g SNAP silicagel cartridge and eluted with a gradient of DCM to 5% MeOH/DCM over 10column volumes. The expected compound was collected and evaporated toyield a tan solid. The tan solid was dissolved in tetrahydrofuran (THF)(10.00 mL) followed by the addition of 1M lithium hydroxide solution (10mL, 10 mmol). The reaction was stirred at 50° C. for 2 h. The reactionwas cooled to room temperature and the organic solvent was removed invacuo. The solution was diluted with water (20 mL) and acidified with 1N HCl. The mixture was then filtered and the grey solid was purified byreversed phase HPLC with a gradient of acetonitrile (0.1% TFA) and water(0.1% TFA v/v) (10-40%) over 10 minutes. The appropriate fractions werecollected and evaporated to provide the desired product (70.5 mg, 0.176mmol, 24.27% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 2.04 (d,J=6.82 Hz, 3H) 2.78 (br. s., 4H) 3.02 (s, 3H) 3.78 (br. s., 4H)5.95-6.06 (m, 1H) 6.49 (br. s., 1H) 7.35-7.51 (m, 3H) 7.57 (m, 1H). MS(ES+) m/e 399.8 [M+H]⁺.

Example 85

Preparation of1-[(3-chlorophenyl)methyl]-2-methyl-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid

To a solution of methyl2-methyl-5-(4-morpholinyl)-1H-benzimidazole-7-carboxylate, prepared asdescribed in Example 26, step d (0.2 g, 0.726 mmol) inN,N-Dimethylformamide (DMF) (10 mL) was added1-(bromomethyl)-3-chlorobenzene (0.190 mL, 1.453 mmol) and potassiumcarbonate (0.301 g, 2.179 mmol). The resulting reaction mixture wasstirred for 3 h at 80° C. The solution was cooled to room temperatureand poured into water and was extracted with EtOAc. The combined organicphase was washed with Brine and concentrated. The residue was purifiedon Biotage Isolera purification system using a Biotage 10 g SNAP silicagel cartridge and eluted with a gradient of DCM to 5% MeOH/DCM over 10column volumes. The expected compound was collected and evaporated toyield a tan solid. The tan solid was dissolved in tetrahydrofuran (THF)(10.00 mL) followed by the addition of 1M lithium hydroxide solution (10mL, 10 mmol). The reaction was stirred at 50° C. for 2 h. The reactionwas cooled to room temperature and the organic solvent was removed invacuo. The solution was diluted with water (20 mL) and acidified with 1N HCl. The mixture was then filtered and a gray solid was isolated. Theaqueous layer was found to contain a significant amount of product andwas evaporated. Both solid and residue were purified by reversed phaseHPLC with a gradient of acetonitrile (0.1% TFA) and water (0.1% TFA v/v)(5-50%) over 10 minutes. The appropriate fractions were collected andevaporated to procide the desired product (37.5 mg, 0.097 mmol, 13.38%yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 2.93 (br. s., 3H) 3.05 (br.s., 4H) 3.77-3.85 (m, 4H) 5.55 (br. s., 2H) 6.88 (s, 1H) 7.09 (br. s.,1H) 7.20 (s, 1H) 7.31-7.39 (m, 3H). MS (ES+) m/e 385.8 [M+H]⁺.

Example 86

Preparation of2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid 2-amino-2-(hydroxymethyl)-1,3-propanediol Salt

Seed Crystal Preparation—Batch 1:

To the2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid (52.9 mg, 0.122 mmol), methanol (2.0 mL) was added. To the slurry,tromethamine(2-amino-2-(hydroxymethyl)-1,3-propanediol) (3.0 M solutionin water, 1.0 equivalent) was added. The slurry was heated to 60 C. andkept stirring at 60 C for 3 hours. The slurry was then cooled slowly(0.1 C/min) to 20 C. Once the temperature of the slurry reached 20 C,the slurry was kept stirring at 20 C for 8 hours. The crystalline solidswere isolated by vacuum filtration. The yield of the desired salt was57.2 mg (85% yield).

Seed Crystal Preparation—Batch 2:

To the2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid (353.0 mg), methanol (14.0 mL) was added. The slurry was heated to60 C. and tromethamine (3.0 M solution in water, 1.0 equivalent) wasadded in four aliquots over 15 minutes followed by the addition ofcrystalline seeds of crystalline tromethamine salt from batch 1. Theslurry was stirred at 60 C for 3 hours, cooled (1 C/min) to 20 C, andstirred at 20 C for 8 hours. The solids were isolated by vacuumfiltration, dried at 60 C under vacuum for 5 hours. The yield of thetromethamine salt was 401.5 mg (˜88.9% yield).

Batch 3:

2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid (40.0 g, 92 mmol) was suspended in Methanol (1.6 L) in a 3 Lrounded-bottom flask. The resulting slurry was heated to 60° C. mixingon a buchii rotary evaporator water bath andtris(hydroxymethyl)aminomethane (3M solution in water) (0.031 L, 92mmol) was added in four aliquots over 15 minutes followed by theaddition of seed crystals as produced by method analogous to Example 86,Batch 2, above (108 mg). This slurry was stirred (flask rotated onbuchii rotovap) at 60° C. for 3 hours, then cooled (˜1° C./min) to 20°C. (room temperature), then finally magnetically stirred at 20° C. (roomtemperature) for 8 hours. The resulting white solid was isolated byvacuum filtration, dried under vacuum at 60° C. for 8 hours to provide2-methyl-1-{[2-methyl-3-(trifluoromethyl)phenyl]methyl}-6-(4-morpholinyl)-1H-benzimidazole-4-carboxylicacid-2-amino-2-(hydroxymethyl)-1,3-propanediol (1:1) (47.76 g, 86 mmol,93% yield) as a white solid. Both proton NMR and LCMS are consistentwith the proposed structure. ¹H NMR (400 MHz, DMSO-d6) δ ppm 7.61 (d,J=7.83 Hz, 1H) 7.37 (d, J=2.27 Hz, 1H) 7.17-7.33 (m, 2H) 6.33 (d, J=7.83Hz, 1H) 5.59 (s, 2H) 3.66-3.80 (m, 4H) 2.98-3.15 (m, 4H) 2.50-2.58 (m,10H) 2.43 (s, 3H); LCMS m/z MH+=434.3.

Example 87

Preparation of1-(3-(chloromethyl)-2-methylbenzyl)-2-methyl-6-morpholino-1H-benzo[d]imidazole-4-carboxylicacid, 2-amino-2-(hydroxymethyl)-1,3-propanediol Salt

1-(3-chloro-2-methylbenzyl)-2-methyl-6-morpholino-1H-benzo[d]imidazole-4-carboxylicacid (10 g, 25.01 mmol) was suspended in Methanol (400 mL) in a 1 Lrounded-bottom flask. The resulting slurry was heated to 60° C. using aBuchii rotary evaporator water bath (no vacuum) andtris(hydroxymethyl)aminomethane (3M solution in water) (8.34 mL, 25.01mmol) was added in four aliquots over 15 minutes. This slurry wasstirred (flask rotated on Buchii rotovap) at 60° C. for 3 hours, thencooled (˜1° C./min) to 20° C. (room temperature), then finallymagnetically stirred at 20° C. (room temperature) for 15 hours. Theresulting white solid was isolated by vacuum filtration, dried undervacuum at 65° C. for 18 hours to provide1-(3-chloro-2-methylbenzyl)-2-methyl-6-morpholino-1H-benzo[d]imidazole-4-carboxylicacid, 2-Amino-2-(hydroxymethyl)-1,3-propanediol salt (11.1 g, 21.09mmol, 84% yield) as a white solid. MS (ES+) m/e: 400.0, 402.0 [M+H]+. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 7.32-7.39 (m, 2H) 7.16 (d, J=2.27 Hz, 1H)7.05 (t, J=7.96 Hz, 1H) 6.05 (d, J=7.58 Hz, 1H) 5.52 (s, 2H) 3.68-3.77(m, 4H) 3.36 (s, 6H) 3.02-3.11 (m, 4H) 2.47 (s, 3H) 2.42 (s, 3H).

Biological Assays

Compounds of the present invention were tested according to thefollowing assays and found as inhibitors of PI3 kinases, particularlyPI3Kβ. The activities (IC₅₀) of exemplified compounds range from about 1nM to about 10 μM against PI3Kβ. The majority of the compounds wereunder 500 nM; the most active compounds were under 10 nM. The IC₅₀ valuecan be converted and presented as pIC₅₀ value.

HTRF In Vitro Profiling Assays for PI3K Inhibition

The PI3-Kinase profiling assays were developed to measure thecompound-dependent inhibition of the alpha, beta, delta, and gammaisoforms of PI3K in an in vitro catalytic assay. This assay wasdeveloped and optimized from a kit produced by Upstate (Milliporecatalog #33-017). Briefly, this procedure utilizes a pre-formed HTRF(Homogeneous Time-Resolved Fluorescence energy transfer) complex betweenfour binding partners: 1) biotinylated PIP3, 2) GST tagged pleckstrinhomology (PH) domain, 3) Europium labeled anti-GST monoclonal antibody,and 4) Streptavidin-Allophycocyanin (APC). The native PIP3 produced byPI 3-Kinase activity displaces biotin-PIP3 from the PH domain, resultingin the dissociation of the HTRF complex and a decrease in thefluorescence signal. The format of this assay is the same for all 4isoforms of PI3K; the differences lie in the concentration of enzymeused to achieve the most robust signal. The alpha and delta assays arerun at 400 μM enzyme; the beta assay is at 200 μM enzyme and the gammaassay is run at 1 nM enzyme. In addition, the alpha, beta and deltaassays are run with 150 mM NaCl while the gamma assay is run in theabsence of NaCl. The ATP concentration is 100 uM in the alpha, beta, anddelta assays and 15 uM ATP in the gamma assay. All reactions are run at10 uM PIP2

Compounds were serially diluted (3-fold in 100% DMSO) across a 384-wellpolypropylene mother plate from column 1 to column 12 and column 13 tocolumn 24, to yield 11 concentrations for each test compound. Columns 6and 18 contain only DMSO. Once titrations were made, 0.05 μL wastransferred to a 384-well low-volume assay plate (Greiner 784076). Thisassay plate contained three pharmacological controls (known PI3Kinhibitors) and 3 assay controls: (1) Enzyme without inhibitor; (2)Buffer minus enzyme, and (3) Buffer minus enzyme plus native PIP3. DMSOwas stamped into all wells of columns 6 and 18. PIP3 was added at 40 μMin 1× Reaction buffer (1 μL of 200 μM PIP3) to alternating rows ofcolumn 18 (wells 18 B, D, F, H, J, L, N, P). The no-enzyme controlreactions were run in wells 18 A, C, E, G, I, K, M, O (0.1 μL of 100%DMSO).

The PI3-Kinase profiling assay was optimized using the HTRF kit providedby Upstate (Millipore). The assay kit contained seven reagents: 1) 4×Reaction Buffer; 2) native PIP2 (substrate); 3) Stop A (EDTA); 4) Stop B(Biotin-PIP3); 5) Detection Mix A (Streptavidin-APC); 6) Detection Mix B(Eu-labeled Anti-GST plus GST-tagged PH-domain); 7) Detection Mix C(KF). In addition, the following items were obtained or purchased:PI3Kinase (prepared by GSK BR&AD), dithiothreitol (Sigma, D-5545),Adenosine-5′-triphosphate (ATP, Teknova cat. # A0220), native PIP3(1,2-dioctanoyl-sn-glycero-3-[phosphoinositil-3,4,5-triphosphate]tetraammoniumsalt (Avanti polar lipids, 850186P), DMSO (Sigma, 472301).

PI3Kinase Reaction Buffer was prepared by diluting the stock 1:4 withde-ionized water. Freshly prepared DTT was added at a finalconcentration of 5 mM on the day of use. Enzyme addition and compoundpre-incubation were initiated by the addition of 2.5 μL of PI3K (attwice its final concentration) in 1× reaction buffer to all wells usinga Multidrop Combi. Plates were incubated at room temperature for 15minutes. Reactions were initiated by addition of 2.5 μL of 2× substratesolution (PIP2 and ATP in lx reaction buffer) using a Multidrop Combi.Plates were incubated at room temperature for one hour. Reactions werequenched by the addition of 2.5 μL of stop solution (Stop A and Stop Bpre-mixed at a ratio of 5:1, respectively) to all wells using theMultidrop Combi. The quenched reactions were then processed to detectproduct formation by adding 2.5 μL of Detection Solution to all wellsusing the Mulitdrop Combi (Detection mix C, Detection mix A, andDetection mix B combined together in an 18:1:1 ratio, i.e.: for a 6000μL total volume, mix 5400 μL Detection mix C, 300 μL Detection mix A,and 300 μL Detection mix B. Note: this solution should be prepared 2hours prior to use). Following a one hour incubation in the dark, theHTRF signal was measured on the Envision plate reader set for 330 nmexcitation and dual emission detection at 620 nm (Eu) and 665 nm (APC).

The loss of the HTRF signal is due to the displacement ofbiotinylated-PIP3 from the PH domain by the PI3K-dependent conversion ofPIP2 to PIP3. This loss of signal is nonlinear with respect to bothincreasing product and time. This non-linear detection will impactaccuracy of IC₅₀ calculations; therefore, there is a need for acorrection factor to obtain more accurate IC₅₀ values. This correctionis derived from the assay standards in the wells of column 6 and 18 ofthe assay plate.

All data were calculated using the ratio of acceptor (APC) to donor(Europium) fluorescence in each well of the assay plate. The percentinhibition for each compound concentration was calculated as follows: %inhibition=100*(fluorescence ratio−CtrlB)/(CtrlA−CtrlB) where CtrlA=(−)PI3Kinase reaction and CrtlB=PI3Kinase+DMSO.

An IC₅₀ was then calculated fitting the % inhibition data to theequation: % inhibition=min+(max−min)/(1+([inhibitor]/IC₅₀)^n) where minis the % inhibition with no inhibitor (typically 0%), max is the signalin the (−) Enzyme control, and n is the Hill slope (typically 1).Finally, the IC₅₀ was converted to pIC₅₀ (pIC₅₀=−log(IC₅₀)), and thepIC₅₀ value was corrected by using plate controls and the equationbelow:

pIC₅₀ (corrected)=pIC₅₀ (observed)+log 10((CtrlA−CtrlB)/(CtrlB−CtrlC)),where CtrlA and CtrlB are as defined above and CrtlC=10 μM PI(3,4,5)P3,100% displacement of biotinylated PI(3,4,5)P3.

The compounds listed in Table 1 were tested generally according to theassays described herein. Table 1 lists the pIC50 values for either anexperimental run or an average of two or more experimental runs with theexamples shown.

TABLE 1 Example # MW PI3KB PIC50 MEAN  3 414.51 6.7  5 373.46 8.8 11401.51 8.7 13 420.34 8.1 14 406.32 8.5 15 375.45 9.0 17 389.48 8.6 18408.31 8.0 20 401.47 8.2 22 420.30 7.8 25 424.51 9.6 31 433.43 8.2 32401.30 7.9 35 402.46 7.2 38 420.30 7.0 39 456.48 9.2 41 399.50 8.8 43419.45 8.3 50 455.44 8.9 53 453.85 9.3 54 486.42 8.6 58 425.50 8.2 59399.88 8.2 63 468.32 9.0 70 469.42 9.7 72 435.86 8.8 73 407.50 7.6Cellular Assays—Cell Growth Inhibition Assay in PTEN Wild-Type or PTENDeficient Tumor Cell Lines

Twenty-two Phosphatase and Tensin Homolog (PTEN) wild-type or PTENdeficient tumor cell lines were cultured generally according toinstructions supplied by cell culture supplier American Type CultureCollection, Manassas, Va., with 10% fetal bovine serum at 5% CO₂ and 37°C. Cells were seeded into either a T-75 or a T-175 flask 3-4 days priorto 96-well assay plating such that the flasks were approximately 70-80%confluent of the time of harvest. Cells were harvested using 0.25%trypsin-EDTA (Invitrogen #25200056). Trypan Blue exclusion staining wasused to determine cell number.

Viable cells were plated in clear, flat bottom 96-well plates (BD#353075) under anchorage independent conditions at 2,000-10,000 cellsper well depending on the cell line. To generate anchorage independentgrowth conditions, a 5% agar stock solution in water was made andautoclaved to melt and sterilize. From the 5% agar solution, a 0.6%agar/media+10% fetal bovine serum (FBS) solution was made to generate abottom agar layer in the plates to prevent cell attachment. Seventy fivemicroliters per well of the 0.6% agar-media solution was added to theplates. After solidification, a cell solution of 266,870 to 1,334,022cells (depending on the cell line) in 10 ml of 0.3% agar/media+10% FBSwas made and 75 μl of the cell/media/agar suspension was added to theplates. After the cell layer solidified, 50 μl of media+10% FBS wasadded to the top of the cells. A 0.3% Brij 35 (Sigma B4184) solution inmedia+10% FBS was added to column 12 as a background subtractioncontrol. The cells were incubated overnight at 5% CO₂ and 37° C. Thenext day one plate of cells was processed at the time of compoundaddition to quantify the starting number of cells (T=0 or T0).

To generate the compound titration plates, 15 μl of a 2 mM or 20 μl of a20 mM solution of the compound of example 31 was diluted in clear bottompolypropylene 96-well plate (BD #351190) using a 10 point, 3-foldtitration or a 20 point 2-fold titration, respectively. Three hundredmicroliters of media was added to the compound dilutions. Tenmicroliters per well of the serial dilutions was added to the cells andthe plates incubated for 6 days at 5% CO₂ and 37° C. The finalconcentration of DMSO in all wells was 0.15% and the highest finalconcentration of the compound of example 31 was 3.7 μM or 30.7 μM.

Following the 6-day incubation, 20 μl of Alamar Blue (Invitrogen#DAL1100) was added to the cells, incubated at 5% CO₂ and 37° C. for 6hours and the plates read on a Spectramax (Gemini EM) at 530 nm(excitation) and 590 nm (emission) with the auto cut-off disabled. Foranalysis of cell growth inhibition dose response curves, the data wasplotted as the percent of the DMSO-treated control samples (DMSO samplesset to 100%). The cellular response was determined for the compound ofexample 31 and control compounds by fitting the concentration responsewith a 4 parameter curve fit using XLfit software and determining theconcentration that inhibits 50% of the Ymax−Ymin window (EC₅₀). The EC₅₀is the midpoint of active compound effect window (between Ymax plateauand Ymin plateau of compound) and represents the concentration of thecompound of example 31 where 50% of its maximal effect is observed.Values from wells containing 0.3% Brij 35 (under anchorage independentconditions) were subtracted from all samples for background correction.

The results shown in Table 2 demonstrate that multiple cell lines withloss of the tumor suppressor PTEN were sensitive, while relatively fewwild-type PTEN tumor cell lines were sensitive.

TABLE 2 Anchorage Independent Soft Agar Tumor Growth Assay PTEN CompoundPTEN Mutation/Copy Western Cell line Origin Type Number Status AnalysisEC₅₀ (nM) ± StDev Ymin ± StDev BT549 breast carcinoma p.V275fs*1 Noprotein  7 ± 2 52 ± 7 WM-115 skin melanoma p.165fs* No protein  8 ± 3 54 ± 14 C32 skin melanoma p.55fs* No protein  8 ± 2  20 ± 14 SW1783 CNSglioblastoma p.R233* No protein 10 ± 3 69 ± 4 UM-UC-3 bladdertransitional Loss No protein 10 ± 8  83 ± 29 SW1088 CNS glioblastomaLoss No protein 12 ± 6 36 ± 8 H4 CNS glioblastoma Loss No protein 12 ± 8 55 ± 18 CHL-1 skin melanoma Wild-type Protein 14 ± 6 82 ± 2 UACC-62skin melanoma p.P248fs*5 No protein  23 ± 27  76 ± 18 HCC19377 breastcarcinoma Loss No protein 24 ± 8 67 ± 8 PC-3 prostate carcinoma Loss Noprotein  27 ± 12  82 ± 13 HCC70 breast carcinoma p.F90fs*9 No protein 53 ± 21 27 ± 8 MDA-MB-468 breast carcinoma p.?, L70Fs*7 No protein  89± 55 44 ± 6 HCC1395 breast carcinoma p.N212fs*3 No protein 114 ± 53 26 ±9 U-87MG CNS glioblastoma p.? No protein  2975 ± 2771  34 ± 18 B1474breast carcinoma Wild-type Protein 3360 ± 868 75 ± 3 U251 CNSglioblastoma p.E242fs*15 No protein 18996 ± 8625  80 ± 17 HCC1954 breastcarcinoma Wild-type Protein >30722 >80 Co1o205 colon carcinoma Wild-typeND >30722 >80 HCT-116 colon carcinoma Wild-type ND >30722 >80 SKOV-3ovary adenocarcinoma Wild-type ND >30722 >80 LOXIMVI skin melanomaWild-type Protein >30722 >90 p.? indicates a splice site mutationIn Vivo ExperimentsDose Dependent Tumor Inhibition

The activity of the compound of example 31 was evaluated in vivo againstPC-3 (prostate carcinoma cell line encoding a deficient PTEN protein)xenograft mouse model. The PC-3 tumor bearing mice were generated byinjecting 2.5×10⁶ PC-3 cells suspended 1:1 in Matrigel subcutaneously inthe flank of female nude mice (Charles River—Wilmington; strain Crl:CD-1-Foxn1). One set of mice, each approximately 19 weeks of age, wereimplanted with the cells for the 100, 30, and 10 mg/kg doses and anotherset of mice, each approximately 11 weeks of age, were implanted with thecells for the 10, 3, and 1 mg/kg doses.

Mice bearing PC-3 xenografts were randomized into dosing groups of n=8based on tumor volume 29 (100, 30, and 10 mg/kg) or 28 (1, 3, 10 mg/kg)days after tumor cells were implanted. Treatment of mice commenced thenext day and continued for 21 days. Mice received once daily oral gavagewith compound or vehicle at 10 mL/kg.

Tumor growth was measured twice weekly in two dimensions with verniercallipers; the longest dimension was defined as the length (1), and thewidth (w) was measured perpendicular to the length. Tumor volumes (V)were calculated using the following equation: V=(½)lw². Means of thetumor volumes were used to compare treatment groups. Stable disease forthis study is defined as a tumor volume which during the course ofcompound treatment does not substantially increase or decrease but stayssimilar to the volume prior to drug treatment compared to vehicletreated in which the tumor volume continues to increase during thecourse of the study. Tumor growth delay is defined as tumor volume thatis reduced during the course of the compound treatment relative tovehicle treated tumor volume.

The results demonstrated that treatment of female nude mice bearing PC-3prostate xenografts with 10, 30, and 100 mg/kg the compound of example31 for 21 days resulted in stable disease with the 1 and 3 mg/kg dosesresulting in tumor growth delay relative to vehicle during the dosingperiod.

B) Pharmacodynamic Effects

The activity of the compound of example 31 was evaluated in vivo againstPC-3 (prostate carcinoma cell line encoding a deficient PTEN protein)xenograft mouse model. Female nude mice (Charles River Laboratories,Wilmington, Del.; strain CD-1-Foxn1, ˜6 weeks of age) were injectedsubcutaneously with 2 million PC-3 (human prostate carcinoma) cellsmixed 1:1 with Matrigel in the flank. Tumors were allowed to grow forapproximately 5 weeks.

Mice bearing PC-3 xenografts were administered 3 mg/kg of the compoundof example 31 or 10 mg/kg of the compound of example 31 and euthanizedusing carbon dioxide after 1, 2, 4, 6, 8, 10, and 24 hours (n=3mice/treatment/timepoint); an additional 3 mice bearing PC-3 xenograftswere administered vehicle and euthanized after 2 hours. The tumor wasexcised. Half of each tumor was immediately processed by Medicon (BDCatalog #340592) in 1 mL Meso-Scale Discovery (MSD) lysis buffer withprotease inhibitors (Roche complete protease cocktail, cat #04 693 116001) and phosphatase inhibitors (Sigma, cat # P2850 and P-5726) for30-60 seconds and transferred to 1.5 mL Eppendorf tubes. Tubes remainedon wet ice until they were centrifuged for 10 minutes at 4° C. atmaximum speed in a tabletop refrigerated centrifuge.

Tumor lysates were serially diluted in 96-well polypropylene plates onwet ice. Lysates (150 μL) were loaded in row 1; rows 2-12 were loadedwith 75 μL of complete Meso Scale Discovery (MSD) lysis buffer (suppliedin MSD kit; # K15100D-3). Samples were serially diluted 2-fold acrossthe plate by sequential transfer of 75 μL through well 11; row 12contained lysis buffer only. MSD Multi-Spot assay plates (whole celllysate kit: Phospho(ser473), Total AKT Assay, catalog #K15100D-3) wereblocked with 150 μL of 3% Blocker A overnight at 4° C. with shakingbefore being washed 4× with 200 μL MSD Tris wash buffer. Fiftymicroliters of the serially diluted lysates were pipetted onto theblocked MSD plates, covered, and incubated overnight at 4° C. withshaking. Plates were washed with Tris buffer as before. Detectionantibody was added (25 μL/well) at a final concentration of 10 nM in 1mL Blocker A and 2 mLTris wash buffer and incubated for 1 hour at roomtemperature with shaking. Plates were washed as described above, beforethe addition of 150 μL of MSD read buffer and read immediately on a 6000MSD plate reader. All work was performed in accordance withInstitutional Animal Care and Use Committee (IACUC) protocols PA0079 andPA0271.

The non-lysate controls in column 12 were averaged and used asbackground to subtract from all wells. P/T AKT was calculated as shown:(phospho AKT(Ser473) signal)/[(phospho AKT(Ser473) signal)+(total AKTsignal)]. Values from three points in each row of diluted samplesidentified as being in the linear range of detection were averaged torepresent each tumor sample's P/T AKT value. Averages and standarddeviations of the P/T AKT value for each group of 3 mice were determinedPercent inhibition was calculated for each group as follows:100-[(sample P/T AKT value)/(vehicle P/T AKT value)]*100.

The compound of example 31 exhibited dose dependent inhibition of thepharmacodynamic marker pAKT (pAKT/tAKT).

Additional References:

The compounds of the present invention can also be tested to determinetheir inhibitory activity at PI3Kα, PI3Kδ, PI3Kβ and PI3Kγ according tointernational patent publication No. WO2009/039140.

The pharmaceutically active compounds within the scope of this inventionare useful as PI3 Kinase inhibitors in mammals, particularly humans, inneed thereof.

The present invention therefore provides a method of treating diseasesassociated with PI3 kinase inhibition, particularly: autoimmunedisorders, inflammatory diseases, cardiovascular diseases,neurodegenerative diseases, allergy, asthma, pancreatitis, multiorganfailure, kidney diseases, platelet aggregation, cancer, sperm motility,transplantation rejection, graft rejection and lung injuries and otherconditions requiring PI3 kinase modulation/inhibition, which comprisesadministering an effective compound of Formula (I) or a pharmaceuticallyacceptable salt, hydrate, solvate or pro-drug thereof. The compounds ofFormula (I) also provide for a method of treating the above indicateddisease states because of their ability to act as PI3 inhibitors. Thedrug may be administered to a patient in need thereof by anyconventional route of administration, including, but not limited to,intravenous, intramuscular, oral, subcutaneous, intradermal, andparenteral.

Exemplary Capsule Composition

An oral dosage form for administering the present invention is producedby filing a standard two piece hard gelatin capsule with the ingredientsin the proportions shown in Table 3, below.

TABLE 3 INGREDIENTS AMOUNTS Compound of example 1 25 mg Lactose 55 mgTalc 16 mg Magnesium Stearate  4 mgExemplary Injectable Parenteral Composition

An injectable form for administering the present invention is producedby stirring 1.5% by weight of compound of example 1 in 10% by volumepropylene glycol in water.

Exemplary Tablet Composition

The sucrose, calcium sulfate dihydrate and an PI3K inhibitor as shown inTable 4 below, are mixed and granulated in the proportions shown with a10% gelatin solution. The wet granules are screened, dried, mixed withthe starch, talc and stearic acid; screened and compressed into atablet.

TABLE 4 INGREDIENTS AMOUNTS Compound of example 1 20 mg calcium sulfatedehydrate 30 mg Sucrose 4 mg Starch 2 mg Talc 1 mg stearic acid 0.5 mg

While the preferred embodiments of the invention are illustrated by theabove, it is to be understood that the invention is not limited to theprecise instructions herein disclosed and that the right to allmodifications coming within the scope of the following claims isreserved.

What is claimed is:
 1. A process for making the compound of formula (4)

comprising the steps of brominating a 2,6-dinitro aniline to provide a4-bromo-2,6-dinitroaniline, reducing said 4-bromo-2,6-dinitroaniline toprovide a di-amino nitro benzene, and reacting said di-amino nitrobenzene with 2,4-pentanedione to provide the compound of formula (4). 2.The process of claim 1, wherein the step of brominating comprisesbrominating a 2,6-dinitro aniline with bromine in acetic acid.
 3. Theprocess of claim 1, wherein the step of reducing comprises reducing said4-bromo-2,6-dinitroaniline with (NH₄ ⁺)₂S.
 4. The process of claim 1,wherein the reacting of said di-amino nitro benzene with2,4-pentanedione is preformed in the presence of a strong acid at refluxtemperatures in an alcoholic solvent.